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1.
Environ Pollut ; 302: 119062, 2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35231537

RESUMO

Lead is a metal that exists naturally in the Earth's crust and is a ubiquitous environmental contaminant. The alleviation of lead toxicity is important to keep human health under lead exposure. Biosynthesized selenium nanoparticle (SeNPs) and selenium-enriched Lactobacillus rhamnosus SHA113 (Se-LRS) were developed in this study, and their potentials in alleviating lead-induced injury to the liver and intestinal tract were evaluated in mice by oral administration for 4 weeks. As results, oral intake of lead acetate (150 mg/kg body weight per day) caused more than 50 times and 100 times lead accumulation in blood and the liver, respectively. Liver function was seriously damaged by the lead exposure, which is indicated as the significantly increased lipid accumulation in the liver, enhanced markers of liver function injury in serum, and occurrence of oxidative stress in liver tissues. Serious injury in intestinal tract was also found under lead exposure, as shown by the decrease of intestinal microbiota diversity and occurrence of oxidative stress. Except the lead content in blood and the liver were lowered by 52% and 58%, respectively, oral administration of Se-LRS protected all the other lead-induced injury markers to the normal level. By the comparison with the effects of normal L. rhamnosus SHA113 and the SeNPs isolated from Se-LRS, high protective effects of Se-LRS can be explained as the extremely high efficiency to promote lead excretion via feces by forming insoluble mixture. These findings illustrate the developed selenium-enriched L. rhamnosus can efficiently protect the liver and intestinal tract from injury by lead.


Assuntos
Enteropatias , Lactobacillus rhamnosus , Selênio , Animais , Chumbo/toxicidade , Fígado , Camundongos , Selênio/farmacologia
2.
Front Immunol ; 13: 880578, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35572521

RESUMO

The current study was conducted to analyze the functions of blood neutrophils in transition cows and their association with postpartum mastitis risk as indicated by somatic cell counts (SCCs) in milk. Seventy-six healthy Holstein dairy cows were monitored from Week 4 prepartum to Week 4 postpartum. Five dairy cows with low SCCs (38 ± 6.0 × 103/mL) and five with high SCCs (3,753 ± 570.0 × 103/mL) were selected based on milk SCCs during the first three weeks of lactation. At Week 1 pre- and postpartum, serum samples were obtained from each cow to measure neutrophil extracellular trap (NET)-related variables, and blood neutrophils were collected for transcriptome analysis by RNA sequencing. The serum concentration of NETs was significantly higher (P < 0.05) in cows with high SCCs than in cows with low SCCs (36.5 ± 2.92 vs. 18.4 ± 1.73 ng/mL). The transcriptomic analysis revealed that the transcriptome differences in neutrophils between high- and low-SCC cows were mainly in cell cycle-related pathways (42.6%), including the cell cycle, DNA damage, and chromosomal conformation, at Week 1 prepartum. The hub genes of these pathways were mainly involved in both the cell cycle and NETosis. These results indicated that the formation of NETs in the blood of transition dairy cows was different between cows with low and high SCCs, which may be used as a potential indicator for the prognosis of postpartum mastitis risk and management strategies of perinatal dairy cows.

3.
J Exp Bot ; 2022 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-35511088

RESUMO

The accumulation of reducing sugars in cold-stored tubers, known as cold-induced sweetening (CIS), negatively affects potato processing quality. The starch-to-sugar interconversion pathways that are altered in cold-stored CIS tubers have been elucidated, but the mechanism that regulates them remains largely unknown. This study identified a CBF/DREB transcription factor (StTINY3) that enhances CIS resistance by both activating starch biosynthesis and repressing the hydrolysis of sucrose to reducing sugars in detached cold-stored tubers. Silencing StTINY3 in a CIS-resistant genotype decreased CIS resistance, while overexpressing StTINY3 in a CIS-sensitive genotype increased CIS resistance and altering StTINY3 expression was associated with expression changes in starch resynthesis-related genes. We showed first, that overexpressing StTINY3 inhibited sucrose hydrolysis by enhancing invertase inhibitor StInvInh2 expression, and second, that StTINY3 promoted starch resynthesis by upregulating a large subunit of ADP-glucose pyrophosphorylase StAGPaseL3, and the glucose-6-phosphate transporter StG6PT2. We also provide evidence that StTINY3, is a nucleus-localized transcriptional activator that directly binds to the dehydration-responsive element/CRT cis-element in the promoters of StInvInh2 and StAGPaseL3 using electrophoretic mobility shift assays. Taken together, these findings establish that StTINY3 influences CIS resistance in cold-stored tubers by coordinately modulating the starch-to-sugar interconversion pathways and is a good target for improving potato processing quality.

4.
Microbiol Spectr ; : e0018522, 2022 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-35579473

RESUMO

Listeria monocytogenes is a foodborne pathogen that can cause invasive disease with high mortality in immunocompromised individuals and can survive in a variety of food-associated environments for a long time. L. monocytogenes clonal complex (CC) 87 is composed of ST87 and three other STs and has been identified as the most common subgroup associated with both foods and human clinical infections in China. Therefore, the persistence of CC87 L. monocytogenes in food-associated environments poses a significant concern for food safety. In this study, 83 draft genomes of CC87 L. monocytogenes, including 60 newly sequenced genomes, were analyzed with all isolates from our previous surveillance in Zigong, Sichuang, China. Sixty-eight of the studied isolates were isolated from one retail market (M1 market), while the others were from seven other markets (M2-M8 markets) in the same city. Whole-genome multilocus sequence typing (wg-MLST) and the whole-genome single nucleotide polymorphism (wg-SNP) analysis were performed. Three persistent contamination routes were identified in the M1 market, caused by 2 clusters (A and B) and a wgST31 type. Cluster A isolates were associated with the persistent contamination in a raw meat stall (M1-S77), while Cluster B isolates caused a persistent contamination in aquatic foods stalls. Five wgST31 isolates caused persistent contamination in a single aquatic stall (M1-S65). A pLM1686-like plasmid was found in all Cluster A isolates. A novel plasmid, pLM1692, a truncated pLM1686 plasmid without the cadmium, and other heavy metal resistance genes were conserved in all wgST31 isolates. By comparing persistent and putative non-persistent isolates, four genes that were all located in the prophage comK might be associated with persistence. These findings enhanced our understanding of the underlying mechanisms of contamination and assist in formulating targeted strategies for the prevention and control of L. monocytogenes transmission from the food processing chain to humans. IMPORTANCE Contamination of food by Listeria monocytogenes at retail level leads to potential consumption of contaminated food with high risk of human infection. Our previous study found persistent contamination of CC87 L. monocytogenes from a retail market in China through pulsed-field gel electrophoresis and multilocus sequence typing. In this study, whole-genome sequencing was used to obtain the highest resolution inference of the source and reasons for persistent contamination; meat grinders and minced meat were the major reservoir of persistent contamination in meat stalls, whereas fishponds were the major reservoir in seafood stalls, with different L. monocytogenes isolates involved. These isolates carried different properties such as plasmids and prophages, which may have contributed to their ability to survive or adapt to the different environments. Our findings suggest that whole-genome sequencing will be an effective surveillance tool to detect persistent L. monocytogenes contamination in retail food markets and to design new control strategies to improve food safety.

5.
BMC Cancer ; 22(1): 545, 2022 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-35568824

RESUMO

BACKGROUND: Laryngeal squamous cell carcinoma (LSCC) is the most common squamous cell carcinoma. Though significant effort has been focused on molecular pathogenesis, development, and recurrence of LSCC, little is known about its relationship with the immune-related long non-coding RNA (lncRNA) pairs. METHODS: After obtaining the transcriptome profiling data sets and the corresponding clinical characteristics of LSCC patients and normal samples from The Cancer Genome Atlas (TCGA) database, a series of bioinformatic analysis was conducted to select the differently expressed immune-related lncRNAs and build a signature of immune-related lncRNA pairs. Then, the effectiveness of the signature was validated. RESULTS: A total of 111 LSCC patients and 12 normal samples' transcriptome profiling data sets were retrieved from TCGA. 301 differently expressed immune-related lncRNAs were identified and 35,225 lncRNA pairs were established. After univariate Cox analysis, LASSO regression and multivariate Cox analysis, 7 lncRNA pairs were eventually selected to construct a signature. The riskscore was computed using the following formula: Riskscore = 0.95 × (AL133330.1|AC132872.3) + (-1.23) × (LINC01094|LINC02154) + 0.65 × (LINC02575|AC122685.1) + (-1.15) × (MIR9-3HG|LINC01748) + 1.45 × (AC092687.3|SNHG12) + (-0.87) × (AC090204.1|AL158166.1) + 0.64 × (LINC01063|Z82243.1). Patients were classified into the high-risk group (> 1.366) and the low-risk group (< 1.366) according to the cutoff value (1.366), which is based on the 5-year riskscore ROC curve. The survival analysis showed that the low-risk group had a better prognosis (P < 0.001). The riskscore was better than other clinical characteristics in prognostic prediction and the area under the curves (AUCs) for the 1-, 3-, and 5-year survivals were 0.796, 0.946, and 0.895, respectively. Combining age, gender, grade, stage, and riskscore, a nomograph was developed to predict survival probability in LSCC patients. Then, the riskscore was confirmed to be related with the content of tumor-infiltration immune cells and the model could serve as a potential predictor for chemosensitivity. CONCLUSION: We successfully established a more stable signature of 7 immune-related lncRNA pairs, which has demonstrated a better prognostic ability for LSCC patients and may assist clinicians to precisely prescribe chemo drugs.

6.
Cell Biosci ; 12(1): 62, 2022 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-35568915

RESUMO

BACKGROUND: Stem cell senescence is considered as a significant driver of organismal aging. As individuals age, the number of stem cells is declined, and the ability to proliferate and survive is also weakened. It has been reported that metabolism plays an important role in stem cell self-renewal, multilineage differentiation, senescence and fate determination, which has aroused widespread concerns. However, whether metabolism-related genes or signalling pathways are involved in physiological aging remain largely undetermined. RESULTS: In the current study, we showed 868 up-regulated and 2006 down-regulated differentially expressed genes (DEGs) in bone marrow mesenchymal stem cells (MSCs) from old rats in comparison with that from young rats by performing RNA sequence. And DEGs functions and pathways were further selected by function enrichment analysis. The results indicated that the high expression of DEGs might participate in cell differentiation, growth factor binding and etc., while the down-regulated DEGs were majorly enriched in metabolism process, such as the cellular metabolic process and mitochondria. Then, we screened and verified DEGs related to glucose metabolism and investigated the glycolysis levels. We identified that glucose uptake, lactate secretion, ATP production and relative extracellular acidification rates (ECAR) were all diminished in MSCs from old rats. More importantly, we conducted microRNA prediction on the key DEGs of glycolysis to elucidate the potential molecular mechanisms of glucose metabolism affecting MSC senescence. CONCLUSIONS: Our study unravelled the profiles of DEGs in age-associated MSC senescence and their functions and pathways. We also clarified DEGs related to glucose metabolism and down-regulated glycolysis level in age-associated MSC senescence. This study will uncover the metabolic effects on regulating stem cell senescence, and provide novel therapeutic targets for ameliorating age-associated phenotypes.

7.
Ann Transl Med ; 10(7): 410, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35530935

RESUMO

Background: Acute pancreatitis (AP) is characterized by acute onset, rapid development, and poor prognosis. Timely diagnosis and identification of the cause are the key to formulating the clinical program and improving the prognosis. There were several studies on this topic but the results varied. This study systematically evaluated and analyzed reports on the comparison of magnetic resonance imaging (MRI) and computed tomography (CT) for the diagnosis of AP in recent years, providing evidence for clinical diagnosis and treatment. Methods: The databases of PubMed, Web of Science, The Cochrane Library, China National Knowledge Infrastructure (CNKI), and Wanfang Data were searched for literature on MRI and CT in the diagnosis of AP. After evaluating the articles and extracting the data, the software RevMan 5.4 and Stata 16.0 were used for meta-analysis. Results: A total of 9 articles were included in the selection, with a total of 566 patients having undergone diagnosis. Meta-analysis showed that for MRI, the diagnostic sensitivity was 92%, 95% confidence interval (CI): 85% to 96%; specificity was 74%, 95% CI: 50% to 89%; positive likelihood ratio was 3.5, 95% CI: 1.6 to 8.0; negative likelihood ratio was 0.11, 95% CI: 0.05 to 0.24; diagnostic odds ratio (DOR) was 32, 95% CI: 7 to 136; and the area under the curve (AUC) value was 0.93, 95% CI: 0.90 to 0.95. For CT, the diagnostic sensitivity was 73%, 95% CI: 55% to 85%; specificity was 64%, 95% CI: 42% to 82%; positive likelihood ratio was 2.0, 95% CI: 1.1 to 3.6; negative likelihood ratio was 0.43, 95% CI: 0.24 to 0.76; DOR was 5, 95% CI: 2 to 14; and the AUC value was 0.74, 95% CI: 0.70 to 0.78. The AUC value of MRI was significantly greater than CT (Z=3.684, P=0.023). Discussion: In the diagnosis of AP, MRI is more sensitive, specific, and accurate than CT, and can be used as the first choice for the diagnosis of AP.

8.
J Nutr Biochem ; : 109039, 2022 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-35533902

RESUMO

Liver fibrosis is a pathological process as a result of intrahepatic deposition of excessive extracellular matrix. Epithelial-mesenchymal transition (EMT) of hepatocytes and activation of hepatic stellate cells (HSCs) both play important roles in the etiology of liver fibrosis. Here, we found that limonin repressed transforming growth factor-ß1 (TGF-ß)-induced EMT in AML-12 hepatocytes and activation of LX-2 HSCs. In both kinds of cells, limonin suppressed TGF-ß-provoked Smad2/3 C-terminal phosphorylation and subsequent nuclear translocation. Transcription of Smad2/3-downstream genes was in turn reduced. However, limonin exerted few effects on Smad2/3 phosphorylation at linker region. Mechanistically, limonin increased Smad7 at mRNA level in both AML-12 and LX-2 cells. Knockdown of Smad7 abrogated inhibitory effects of limonin on TGF-ß-induced EMT in AML-12 cells and activation of LX-2 cells. Further studies revealed that limonin alleviated mouse liver fibrosis induced by CCl4. In livers of model mice, limonin upregulated Smad7 and declined C-terminal phosphorylation and nuclear translocation of Smad2/3. Transcription of Smad2/3-responsive genes was also attenuated. Our findings indicated that limonin inhibits TGF-ß-induced EMT of hepatocytes and activation of HSCs in vitro and CCl4-induced liver fibrosis in mice. Upregulated Smad7 which suppresses Smad2/3-dependent gene transcription is implicated in the hepatoprotective activity of limonin.

9.
Front Endocrinol (Lausanne) ; 13: 872153, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35527993

RESUMO

BRAFV600E is the most common mutated gene in thyroid cancer and is most closely related to papillary thyroid carcinoma(PTC). We investigated the value of elasticity and grayscale ultrasonography for predicting BRAFV600E mutations in PTC. Methods: 138 patients with PTC who underwent preoperative ultrasound between January 2014 and 2021 were retrospectively examined. Patients were divided into BRAFV600E mutation-free group (n=75) and BRAFV600E mutation group (n=63). Patients were randomly divided into training (n=96) and test (n=42) groups. A total of 479 radiomic features were extracted from the grayscale and elasticity ultra-sonograms. Regression analysis was done to select the features that provided the most information. Then, 10-fold cross-validation was used to compare the performance of different classification algorithms. Logistic regression was used to predict BRAFV600E mutations. Results: Eight radiomics features were extracted from the grayscale ultrasonogram, and five radiomics features were extracted from the elasticity ultrasonogram. Three models were developed using these radiomic features. The models were derived from elasticity ultrasound, grayscale ultrasound, and a combination of grayscale and elasticity ultrasound, with areas under the curve (AUC) 0.952 [95% confidence interval (CI), 0.914-0.990], AUC 0.792 [95% CI, 0.703-0.882], and AUC 0.985 [95% CI, 0.965-1.000] in the training dataset, AUC 0.931 [95% CI, 0.841-1.000], AUC 0. 725 [95% CI, 0.569-0.880], and AUC 0.938 [95% CI, 0.851-1.000] in the test dataset, respectively. Conclusion: The radiomic model based on grayscale and elasticity ultrasound had a good predictive value for BRAFV600E gene mutations in patients with PTC.


Assuntos
Proteínas Proto-Oncogênicas B-raf , Neoplasias da Glândula Tireoide , Elasticidade , Humanos , Mutação , Proteínas Proto-Oncogênicas B-raf/genética , Estudos Retrospectivos , Câncer Papilífero da Tireoide/diagnóstico por imagem , Câncer Papilífero da Tireoide/genética , Câncer Papilífero da Tireoide/patologia , Neoplasias da Glândula Tireoide/diagnóstico por imagem , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/patologia , Ultrassonografia
10.
Artigo em Inglês | MEDLINE | ID: mdl-35522641

RESUMO

The COVID-19 pandemic has spread the world like no other crisis in recent history. Automatic segmentation of COVID-19 pneumonia lesions is critical for quantitative measurement for diagnosis and treatment management. For this task, deep learning is the state-of-the-art method while requires a large set of accurately annotated images for training, which is difficult to obtain due to limited access to experts and the time-consuming annotation process. To address this problem, we aim to train the segmentation network from imperfect annotations, where the training set consists of a small clean set of accurately annotated images by experts and a large noisy set of inaccurate annotations by non-experts. To avoid the labels with different qualities corrupting the segmentation model, we propose a new approach to train segmentation networks to deal with noisy labels. We introduce a dual-branch network to separately learn from the accurate and noisy annotations. To fully exploit the imperfect annotations as well as suppressing the noise, we design a Divergence-Aware Selective Training (DAST) strategy, where a divergence-aware noisiness score is used to identify severely noisy annotations and slightly noisy annotations. For severely noisy samples we use an unsupervised regularization through dual-branch consistency between predictions from the two branches. We also refine slightly noisy samples and use them as supplementary data for the clean branch to avoid overfitting. Experimental results show that our method achieves a higher performance than standard training process for COVID-19 pneumonia lesion segmentation when learning from imperfect labels, and our framework outperforms the state-of-the-art noise-tolerate methods significantly with various clean label percentages.

11.
Plant Dis ; 2022 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-35536209

RESUMO

Magnolia × soulangeana Soul.-Bod., the saucer magnolia is an important woody ornamental plant cultivated widely in China, UK and USA. In August 2021, symptoms and signs of powdery mildew appeared on leaves of M. × soulangeana at the campus of Nanjing Forestry University (NJFU). The powdery mildew mainly infected young seedlings, with an incidence of 96.8% (436/450 seedlings), and some adult trees also been infected (5/30 trees). The mycelium was amphigenous, thinly effused or conspicuous, forming circular to irregular white patches. Noticeable brown lesions and necrosis occurred in the later stage of infection. Chasmothecia started to develop in October, 2021 and fully matured in early November, 2021. Ten fresh specimens were collected and examined to identify of the pathogen. Photos were taken with a ZEISS Axio Imager A2m microscope, a Zeiss stereo microscope (SteRo Discovery v20), and a scanning electronic microscope (JSM-7600F). Conidiophores arose from the upper part of mother cells, 78.5 ± 11.2× 10.9 ± 1.7 µm (n=30). Foot cells in conidiophores are straight and cylindrical with a constricted basal septum close to hyphal mother cell, 33.6 ± 4.3 × 10.3 ± 1.2 µm (n=30). Conidia were hyaline, ellipsoid to oval, solitary or in chains of two to four, 38.5 ± 3.3 × 18.4 ± 1.0 µm (n=30). Chasmothecia were amphigenous, scattered or aggregated, blackish brown, oblate, 101.1 ± 11.4µm diam. (n=30), with 6-10 appendages. Appendages were aseptate, rarely 1-septate, 5-6 times frequently dichotomously branched; tips were noticeably recurved, brown at the base, 105.1 ± 10.7 × 8.5 ± 1.4 µm (n=30). Asci were 6 to 8 per chasmothecium (n=30), ellipsoid to obovoid or saccate with a short stalk or sessile, 64.2 ± 6.5× 46.1 ± 5.7 um (n=30) in length, 4 to 6 spored. Ascospores were oblong-ovoid, 26.2 ± 1.4 × 13.8 ± 0.7 µm (n=30). Based on the morphological characteristics, the fungus was identified as Erysiphe magnoliicola S.E. Cho, S. Takam. & H.D. Shin. To confirm the causal fungus identity, a representative voucher specimen collected and deposited in herbarium of NJFU (NF50000008) was used for a phylogenetic analysis. Mycelia and conidia were collected from diseased leaves and genomic DNA of the pathogen was extracted. The internal transcribed spacer region (ITS) and large subunit (LSU) loci were amplified with primers ITS1/ITS4 and LR0R/LR05. The resulting sequences were deposited in GenBank (OL454094 for ITS, OM758416 for LSU). BLAST results showed that the ITS sequence was highly similar with a sequence of E. magnoliicola (type) [KJ567072, 614/619 (99.2%)], while LSU sequence was highly similar with E. magnoliicola [KJ567068, 889/891 (99.8%)] and E. magnoliae [JX235969, 903/909 (99.3%)]. Phylogenetic analyses using ITS and LSU sequences with maximum likelihood and Bayesian posterior probability using IQ-TREE v. 1.6.8 and MrBayes v. 3.2.6 placed this fungus in the E. magnoliicola clade. Based on the morphology and phylogeny, the fungus was identified as E. magnoliicola. Pathogenicity tests were carried out on six potted plants of M. × soulangeana. Three seedlings were inoculated by gently pressing the naturally infected leaves onto healthy leaves. Healthy leaves from three other seedlings served as control. Inoculated and control seedlings were placed in separate growth chambers at 23 ± 2°C/16 ± 2°C, 70% relative humidity, with a 16 h/8 h light/dark period. Symptoms developed 10 days after inoculation. The powdery mildew developing on the inoculated seedlings was examined, sequenced and confirmed as E. magnoliicola. The control leaves did not develop powdery mildew. Magnolia × soulangeana is a hybrid of Magnolia denudata × Magnolia liliiflora, both species, as well as M. sieboldii were already known as host plants of E. magnoliicola. This is the first report of powdery mildew caused by E. magnoliicola on M. × soulangeana. This finding provides crucial information for developing effective strategies to monitor and manage this disease.

12.
FASEB J ; 36 Suppl 12022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35554417

RESUMO

We used NaV 1.9 knockout (KO) mice to evaluate the role of NaV 1.9 in chemical activation of vagal C-fiber terminals in the mouse lungs. Single-cell rt-PCR of lung-labeled neurons revealed that the vast majority of TRPV1 expressing vagal sensory neurons innervating the lungs express NaV 1.9 mRNA. The mouse lungs were isolated with the vagus nerves and vagal ganglia intact. Action potentials (APs) were recorded using extracellular electrodes positioned near the relevant cell bodies in the vagal ganglion. The stimuli were applied to the receptive field via perfusion through the mouse trachea. The PAR1 agonist, TFLLR(10mM) evoked 270 ± 62 APs in wild type mice (n=23), but only 85 ± 38 action potentials in NaV 1.9 -/- mice (P< 0.01, n=29). PAR1 is a GPCR stimulus, so we next evaluated a,b-methylene ATP (1ml, 1µM) that stimulates nodose C-fibers in the mouse lung via activation of the ionotropic P2X2/3 receptor. a,b-Methylene ATP evoked 93 ± 18 APs in wildtype mice (n=33), compared to only 13 ± 4 APs the NaV 1.9 -/- mice (P=0.0001, n= 35). The C-fibers of both WT and NaV 1.9 -/- were equally activated by rapid punctate mechanical stimulation of the receptive field with Von Frey fibers. At the patched clamped cell soma, there was no difference in active or passive electrophysiological properties of nodose neurons isolated from WT versus KO animals; rheobases were also the same with AP threshold averaging 0.34 ± 0.1 and 0.29 ± 0.06 mV/ms, respectively. However, bath-applied ATP was more effective in evoking action potentials in neurons isolated from WT (40 % responding) vs. KO (0 % responding). The data reveal that NaV 1.9 is important in the activation of nodose C-fiber terminals evoked by inflammatory mediators that act via GPCRs or ionotropic receptors but less critical in the activation by rapid punctate mechanical or electrical stimulation. NaV 1.9 provides an intriguing target for treating symptoms caused by inflammatory mediator-induced C-fiber activation in the airways, e.g. chronic coughing and excessive reflex bronchospasm and secretions associated with inflammatory airway disease.

13.
FASEB J ; 36 Suppl 12022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35556110

RESUMO

Hypertension has now been closely associated to cognitive impairment, due to its contribution to cerebrovascular dysfunction. To investigate whether there is neuronal mechanism, hypertension was induced in C57BL/6j male mice (6 months) by DOCA-salt treatment (1.5 mg/g, s.c., 28 days). Firstly, behavioral study, which contains object location behavioral task, novel object recognition, Barnes maze, and nesting score test, was performed to assess cognitive function, meanwhile, blood pressure (BP) was monitored by tail-cuff device. In NT (non-transgenic) mice, DOCA+1%NaCl increases BP significantly, before vs. after DOCA (systolic BP): 114.8 ±0.94 vs. 133.7 ±3.3 mmHg. In whole, compared to sham controls, the treated NT mice performed worse in all four behavioral tests, indicating cognition decline after DOCA-salt treatment. Later, brain tissues were collected to evaluate possible degenerative alterations in neurons by transmission electron microscope (TEM) imaging, Golgi-Cox staining, fluorescence-activated cell sorting (FACS) and qRT-PCR. It was shown via the TEM that DOCA-salt hypertension induces mitochondrial damage and morphological changes in prefrontal cortical neurons, meanwhile, Golgi-Cox staining revealed decreased numbers of dendritic spines, suggesting the onset of neurodegeneration in DOCA-salt hypertension. In both prefrontal cortex and hippocampus, the mRNA levels of BDNF (brain-derived neurotrophic factor) and PI4KIIIß (phosphatidylinositol 4-kinase IIIß) were found to be significantly lower in hypertensive NT mice. Using FACS, neurons were sorted from those regions, then analyzed via qRT-PCR, which identified that the downregulated expression of PI4KIIIß takes place in neurons. Using laser speckle contrast imaging system, we detected only decreased functional hyperemia, but no impairment in basal cerebral blood flow of the hypertensive mice, suggesting that the neurodegeneration is not caused by cerebral ischemia. We previously demonstrated that neuronal angiotensin type 1 receptors (AT1 R) plays a pivotal role in the maintenance of neurogenic hypertension, and here we hypothesized that activation of AT1 R could also exacerbate hypertension-induced reduction in neuronal function. Particularly, in the hippocampal CA1 of adult mice, treatment of angiotensin II (Ang-II, 500 nM, bath application) induces a significant inhibition in the induction of long-term potentiation (LTP), while pre-treatment with losartan (5 µM, 15 min), the AT1 R antagonist, can efficiently block this Ang-II-induced inhibition, supporting the involvement of neuronal AT1 R. Indeed, in mice with selective deletion of neuronal AT1a R, DOCA-salt-induced neurodegeneration was attenuated, as evidenced by significantly higher mRNA levels of cortical BDNF and PI4KIIIß, compared with the NT. In summary, our data suggest that neuron-expressing AT1 R participates in the development of hypertension-associated cognitive impairment, independently of vascular AT1 R.

14.
FASEB J ; 36 Suppl 12022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35560923

RESUMO

The proper function of airway vagal afferent nerves is essential for the dynamic regulation of breathing and initiation of adequate airway defensive reflexes. A better understanding of the mechanisms controlling their activation bears physiological and pathological significance. The voltage-gated potassium (KV ) channels exert powerful controls on the neuronal excitability. Yet their expression and functions in the bronchopulmonary vagal afferent neurons remain largely unexplored. In this study we characterized the expression profile and inhibitory function of α-dendrotoxin (α-DTX)-sensitive D-type K+ channels composed of members of KV 1subfamily of α-subunits in mouse bronchopulmonary nodose neurons using a combination of single-neurons RT-PCR, patch clamp recording, ex-vivo extracellular recording and two-photon microscopic Ca2+ imaging techniques. The results showed that the vast majority of retrogradely labeled lung-specific nodose neurons expressed Kcna1, 2 and 6 coding for the α-DTX-sensitive KV 1.1, 1.2 and 1.6 α-subunits, respectively. The D-type K+ current (IK.D ), defined as the α-DTX-sensitive K+ current, was recorded in 14/15 bronchopulmonary nodose neurons. It started to be activated at -65.7±4.3 mV with 50% channel activation at -24 ± 14 mV. IK.D opened rapidly in response to depolarization (activation time constants <10 ms at voltage ≥-15 mV), and displayed no inactivation during 600 ms at subthreshold/ threshold voltages and little or slow inactivation at more positive potentials (≥-5 mV). Inhibition of IK.D with 50 nM α-DTX depolarized mouse pulmonary nodose neurons, increased input resistance, lowered the minimal depolarizing current needed to evoke an action potential, and increased the number and frequency of action potential firing in response to depolarizing current injections. Application of a bolus of 100 nM α-DTX to the afferent terminals in the mouse lungs via trachea led to overt activation in 49% of capsaicin-sensitive nodose neurons and in 32% of capsaicin-insensitive nodose neurons as detected by two-photon microscope. Further study with extracellular recording revealed that the application of an α-DTX bolus evoked action potential discharges in 8/19 (42%) nodose C-fibers terminating in the mouse lungs with a peak firing frequency of 4.4 ± 3.9 Hz. These results indicate that both the soma and terminals of bronchopulmonary nodose afferent nerves express functional IK.D characterized by low-threshold, fast activation, and lack of or slow inactivation. With these unique biophysical properties, IK.D channels act as an activation brake on the airway vagal afferent nerves by stabilizing the resting membrane potential and by counterbalancing the subthreshold depolarizing forces. Down-regulation of IK.D , as occurs in many inflammatory diseases, may result in a heightened excitability of airway vagal afferent nerves causing exaggerated dyspnea, excessive mucous secretion, bronchoconstriction and chronic unproductive coughs associated with airway inflammation.

15.
J Adv Res ; 37: 1-18, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35499046

RESUMO

Introduction: Dairy cattle are a vitally important ruminant in meeting the demands for high-quality animal protein production worldwide. The complicated biological process of converting human indigestible biomass into highly digestible and nutritious milk is orchestrated by various tissues. However, poorly understanding of the cellular composition and function of the key metabolic tissues hinders the improvement of health and performance of domestic ruminants. Objectives: The cellular heterogeneity, metabolic features, interactions across ten tissue types of lactating dairy cattle were studied at single-cell resolution in the current study. Methods: Unbiased single-cell RNA-sequencing and analysis were performed on the rumen, reticulum, omasum, abomasum, ileum, rectum, liver, salivary gland, mammary gland, and peripheral blood of lactating dairy cattle. Immunofluorescences and fluorescence in situ hybridization were performed to verify cell identity. Results: In this study, we constructed a single-cell landscape covering 88,013 high-quality (500 < genes < 4,000, UMI < 50, 000, and mitochondrial gene ratio < 40% or 15%) single cells and identified 55 major cell types in lactating dairy cattle. Our systematic survey of the gene expression profiles and metabolic features of epithelial cells related to nutrient transport revealed cell subtypes that have preferential absorption of different nutrients. Importantly, we found that T helper type 17 (Th17) cells (highly expressing CD4 and IL17A) were specifically enriched in the forestomach tissues and predominantly interacted with the epithelial cell subtypes with high potential uptake capacities of short-chain fatty acids through IL-17 signaling. Furthermore, the comparison between IL17RA high IL17RC high cells (epithelial cells with IL17RA and IL17RC expression levels both greater than 0.25) and other cells explained the importance of Th17 cells in regulating the epithelial cellular transcriptional response to nutrient transport in the forestomach. Conclusion: The findings enhance our understanding of the cellular biology of ruminants and open new avenues for improved animal production of dairy cattle.


Assuntos
Lactação , Transcriptoma , Animais , Bovinos , Feminino , Hibridização in Situ Fluorescente , Lactação/fisiologia , Nutrientes , Rúmen
16.
Front Oncol ; 12: 842787, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35574415

RESUMO

Background: Cumulative evidence in colorectal cancer (CRC) suggests that patients with human epidermal growth factor receptor 2 (HER2) overexpression or amplification can benefit from anti-HER2 therapy. The purpose of our study was to evaluate HER2 status and its correlation with clinicopathological characteristics and survival according to currently utilized HER2 diagnostic criteria in a large cohort of Chinese CRC patients. Methods: HER2 protein expression was tested by immunohistochemistry (IHC) in formalin-fixed, paraffin-embedded (FFPE) samples from 4,836 CRC patients in our institution. Breast cancer (BC) and gastroesophageal adenocarcinoma (GEA) criteria, as well as the HERACLES criteria, were used for the determination of HER2 status. Dual-color silver-enhanced in situ hybridization (DSISH) was performed in all IHC 2+~3+ cases determined by BC/GEA criteria. Results: The HER2 expression rate of IHC (1+~3+) was 7.01% (339/4,836) and 6.02% (291/4,836) in CRCs based on the BC/GEA criteria and the HERACLES criteria, respectively, while combined DSISH results in the HER2 amplification/overexpression ratio of 3.39% (164/4,836) in our cohort. HER2 expression detected by IHC was positively correlated with the female gender, whereas the HER2 overexpression/amplification showed no correlation with any clinicopathological parameter. In addition, no significant correlation was found between HER2 statuses and either disease-free survival or overall survival regardless of the evaluation criterion used. However, patients with HER2 1+ CRC showed a tendency of having the shortest overall survival as compared with any other group of patients according to the HERACLES criteria, and this trend has always existed in the rectal location, T3 stage, and TNM stage II, medium differentiation, and perineural invasion stratified group. Furthermore, the HER2 protein expression was significantly negatively correlated with RAS/BRAF mutations according to the HERACLES criteria. Conclusion: To our knowledge, this is the largest study of HER2 status in Asian patients with CRC. Our findings suggest that the current most commonly used HERACLES criteria might be too strict for patients with CRC. Future studies are needed to explore the most suitable criteria for screening CRC patients who could benefit from anti-HER2 therapy as much as possible.

17.
Burns ; 2022 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-35525770
18.
Phytomedicine ; 101: 154087, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35429924

RESUMO

BACKGROUND: Although triple-negative breast cancer (TNBC) accounts for only 15% of breast cancer cases, it is associated with a high relapse rate and poor outcome after standard treatment. Currently, the effective drugs and treatment strategies for TNBC remain limited, and thus, developing effective treatments for TNBC is pressing. Several studies have demonstrated that both chalcone and syringaldehyde have anticancer effect, but their potential anti-TNBC bioactivity are still unknown. PURPOSE: The present study aimed to synthesize a chalcone-syringaldehyde hybrid (CSH1) and explore its potential anti-TNBC effects and the underlying molecular mechanism. METHODS: Cell cytotoxicity was determined by 3-(4,5-dimethythiazol)-2,5-diphenyltetrazolium bromide (MTT). The activity of cell proliferation was measured by colony formation assay and 5-ethynyl-2'-deoxyuridine (EdU) staining assay. Cell cycle distribution and cell apoptosis were determined by fluorescence-activated cell sorter (FACS). The situation of DNA damage was observed using fluorescence microscopy. The ability of cell-matrix adhesion, migration and invasion was detected using cell adhesion assay and transwell assay. Transcriptome sequencing was performed to find out the changed genes. Levels of various signaling proteins were assessed by western blotting. RESULTS: CSH1 treatment triggered DNA damage and inhibited DNA replication, cell cycle arrest, and cell apoptosis via suppressing signal transducer and activator of transcription 3 (STAT3) phosphorylation. Whole genome RNA-seq analysis suggested that 4% of changed genes were correlated to DNA damage and repair, and nearly 18% of changed genes were functionally related to cell adhesion and migration. Experimental evidence indicated that CSH1 treatment significantly affected the distribution of focal adhesion kinase (FAK) and its phosphorylation, resulting in cell-matrix-adhesion reduction and migration inhibition of TNBC cells. Further mechanistic studies indicated that CSH1 inhibited TNBC cell proliferation, adhesion, and migration by inhibiting cytoskeleton-associated protein 2 (CKAP2)-mediated FAK and STAT3 phosphorylation signaling. CONCLUSION: These results suggest that CKAP2-mediated FAK and STAT3 phosphorylation signaling is a valuable target for TNBC treatment, and these findings also reveal the potential of CSH1 as a prospective TNBC drug.

19.
Int J Mol Sci ; 23(8)2022 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-35457035

RESUMO

Osteocytes function as the master orchestrator of bone remodeling activity in the telophase of osseointegration. However, most contemporary studies focus on the manipulation of osteoblast and/or osteoclast functionality via implant surface engineering, which neglects the pivotal role of osteocytes in de novo bone formation. It is confirmative that osteocyte processes extend directly to the implant surface, but whether the surface physicochemical properties can affect the functionality of osteocytes and determine the fate of the osseointegration in the final remodeling stage remains to be determined. Titania nanotube arrays (NTAs) with distinct diameters were fabricated to investigate the relationship between the nanoscale topography and the functionality of osteocytes. In vitro results pinpointed that NTA with a diameter of 15 nm (NTA-15) significantly promote osteogenesis of osteocytes via the enhancement of spreading, proliferation, and mineralization. The osteocyte transcriptome of each group further revealed that the TGF-ß signaling pathway plays a pivotal role in osteocyte-mediated osteogenesis. The in vivo study definitely mirrored the aforementioned results, that NTA-15 significantly promotes bone formation around the implant. Consequently, nanoscaled topography-induced osteocyte functionality is important in late osseointegration. This suggests that surface designs targeting osteocytes may, therefore, be a potential approach to solving the aseptic loosening of the implant, and thus strengthen osseointegration.


Assuntos
Osteócitos , Osteogênese , Osteoblastos/metabolismo , Osteócitos/metabolismo , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo
20.
Stem Cell Rev Rep ; 2022 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-35460064

RESUMO

Cellular senescence is an irreversible cell arrest process, which is determined by a variety of complicated mechanisms, including telomere attrition, mitochondrial dysfunction, metabolic disorders, loss of protein homeostasis, epigenetic changes, etc. Cellular senescence is causally related to the occurrence and development of age-related disease. The elderly is liable to suffer from disorders such as neurodegenerative diseases, cancer, and diabetes. Therefore, it is increasingly imperative to explore specific countermeasures for the treatment of age-related diseases. Numerous studies on humans and mice emphasize the significance of metabolic imbalance caused by short telomeres and mitochondrial damages in the onset of age-related diseases. Although the experimental data are relatively independent, more and more evidences have shown that there is mutual crosstalk between telomeres and mitochondrial metabolism in the process of cellular senescence. This review systematically discusses the relationship between telomere length, mitochondrial metabolic disorder, as well as their underlying mechanisms for cellular senescence and age-related diseases. Future studies on telomere and mitochondrial metabolism may shed light on potential therapeutic strategies for age-related diseases. Graphical Abstract The characteristics of cellular senescence mainly include mitochondrial dysfunction and telomere attrition. Mitochondrial dysfunction will cause mitochondrial metabolic disorders, including decreased ATP production, increased ROS production, as well as enhanced cellular apoptosis. While oxidative stress reaction to produce ROS, leads to DNA damage, and eventually influences telomere length. Under the stimulation of oxidative stress, telomerase catalytic subunit TERT mainly plays an inhibitory role on oxidative stress, reduces the production of ROS and protects telomere function. Concurrently, mitochondrial dysfunction and telomere attrition eventually induce a range of age-related diseases, such as T2DM, osteoporosis, AD, etc. :increase; :reduce;⟝:inhibition.

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