Epithelial-mesenchymal transition (EMT) is an essential component of metastasis. Our previous study demonstrated that cancer-associated fibroblasts (CAFs) induce EMT in lung cancer cells. In recent ...years, many studies have demonstrated that CAFs induce metastasis and drug resistance in cancer cells via exosomes.
We sought to discover the mechanism underlying how CAFs induce EMT in lung cancer cells, unveiling the role of exosomes in lung cancer progression.
We cultured lung cancer cell (i) with control medium, normal fibroblasts (NFs) or CAFs; (ii) with SNAI1-transfected or NC (negative control)-transfected CAFs; (iii) with exosomes extracted from NF- or CAF-conditioned medium; (iv) with exosomes released by SNAI1 or NC-transfected CAFs; (v) with CAF-conditioned medium or exosome-depleted CAF-conditioned medium.
qRT-PCR was conducted to examine the expression of CDH1 (gene of E-cadherin) and VIM (gene of Vimentin), western blotting was conducted to examine E-cadherin and vimentin levels in lung cancer cells.
Exosomes released by CAFs-promoted EMT in lung cancer cells. Interestingly, SNAI1 levels in exosomes secreted from CAFs were correlated with SNAI1 expression in CAFs. Furthermore, the level of SNAI1 in exosomes was crucial for inducing EMT in lung cancer cells. Finally, treatment of CAFs with GW4869, an inhibitor of exosome release, noticeably inhibited their EMT-inducing effect on recipient epithelial cells.
The molecular mechanism underlying how CAFs induce EMT in cancer cells may be that CAFs deliver SNAI1 to recipient cancer cells via exosomes.
Aim
Irisin, a novel myocyte‐secreted hormone mediating beneficial effects of exercise on metabolism, is supposed to be an ideal therapeutic target for metabolic disorders such as obesity and ...diabetes. Here, we investigated the potential effects of metformin and glibenclamide, two antidiabetic medicines, on irisin release in mouse.
Methods
Wild‐type and diabetic obese db/db mice were administrated with metformin and glibenclamide for 2 weeks, and cultured C2C12 myotubes were treated by metformin. Expression of irisin precursor FNDC5 was measured and blood irisin concentration was detected. AMP‐activated protein kinase (AMPK) was blocked by chemical inhibitor compound C or knocking down with specific siRNA.
Results
The mRNA and protein expression of FNDC5 in skeletal muscle and blood irisin concentrations were lower in diabetic db/db mice than those in wild‐type mice. Metformin and glibenclamide decreased blood glucose in db/db mice. Metformin, but not glibenclamide, increased intramuscular FNDC5 mRNA/protein expression and blood irisin levels. Additionally, the reductions of blood glucose and body weight in metformin‐treated db/db mice were positively associated with blood irisin concentrations. In C2C12 myotubes, metformin upregulated intracellular FDNC5 mRNA/protein expression and promoted irisin release. Although metformin activated AMPK signalling in skeletal muscle cells, disrupting of AMPK signalling by chemical inhibitor or siRNA‐mediated knockdown did not abolish the promoting effect of metformin on irisin release.
Conclusion
Metformin promotes irisin release from murine skeletal muscle into blood, independently of AMPK pathway activation. Our results suggest that stimulation of irisin may be a novel molecular mechanism of metformin which is widely used for treatment of metabolic disorders.
Companion animals play an important role in our lives and are now considered to be and treated as family members in a majority of households in the United States. Because of the high number of pets ...that now exist, an increasingly stronger pet-human bond, and the importance placed on health and longevity, the pet food industry has realized steady growth over the last few decades. Despite past successes and opportunities that exist in the future, there are also challenges that must be considered. This review will present a brief overview of the current pet food industry and address some of the key issues moving forward. In regards to companion animal research, recent advances and future needs in the areas of canine and feline metabolism, aging, clinical disease, and the gut microbiome using molecular and high-throughput assays; chemical, in vitro, and in vivo testing of feed ingredients; and innovative pet food processing methods is discussed. Training the future workforce for the pet food industry is also of great importance. Recent trends on student demographics and their species and careers of interest, changing animal science department curricula, and technology's impact on instruction are provided. Finally, the sustainability of the pet food industry is discussed. Focus was primarily placed on the disconnect that exists between opinions and trends of consumers and the nutrient recommendations for dogs and cats, the desire for increasing use of animal-based and human-grade products, the overfeeding of pets and the pet obesity crisis, and the issues that involve the evaluation of primary vs. secondary products in terms of sustainability. Moving forward, the pet food industry will need to anticipate and address challenges that arise, especially those pertaining to consumer expectations, the regulatory environment, and sustainability. Given the already strong and increasingly dynamic market for pet foods and supplies, an academic environment primed to supply a skilled workforce, and continued industry support for basic and applied research initiatives, the future of the pet food industry looks very bright.
In this study, the complete foxl2 complementary (c)DNA sequence was isolated by simple modular‐architecture research tool (SMART)er rapid amplification of cDNA ends (RACE). Two year‐old female ...spotted scat, Scatophagus argus, were reared at different temperatures (23, 26 and 29° C) for 6 weeks, or fed with different concentrations of dietary fish oil (0, 2 or 6%) for 8 weeks. Ovarian development, serum oestradiol‐17β (E2) levels, as well as ovarian foxl2 expression were measured. At the end of experiment, ovarian foxl2 messenger (m)RNA expression in fish reared at 23 and 26° C was significantly higher than that in fish reared at 29° C, and that in 2 and 6% fish oil groups was also significantly higher than that in control group (P < 0·05). Serum E2 levels exhibited the same trend with foxl2 mRNA expression in temperature treatment groups and fish oil fed groups. There was a significant positive correlation between stage of oocytes and foxl2 expressions. Results showed that from 23 to 29° C, the optimal temperature for ovarian development in S. argus was 23–26° C, and 6% fish oil supplementation could effectively promote ovarian development. Optimal temperature and fish oil supplement might increase ovarian foxl2 mRNA expressions to promote ovarian development in S. argus.
Background. Previous studies have shown that common human coronavirus might be neurotropic, although it was first isolated as a pathogen of the respiratory tract. We noticed that a few patients with ...severe acute respiratory syndrome (SARS) experienced central nervous symptoms during the course of illness. In the present study, we isolated a SARS coronavirus strain from a brain tissue specimen obtained from a patient with SARS with significant central nervous symptoms. Methods. Using transmission electronic microscopy and nested reverse transcription–polymerase chain reaction, the causative pathogen was identified in cultures of a brain tissue specimen obtained from the patient with SARS. Histopathologic examination of the brain tissue was performed using the methods of immunohistochemistry analysis and double immunofluorescence staining. Fifteen cytokines and chemokines were detected in the blood of the patient with SARS by means of a bead-based multiassay system. Results. A fragment specific for SARS human coronavirus was amplified from cultures of the brain suspension, and transmission electronic microscopy revealed the presence of an enveloped virus morphologically compatible with a coronavirus isolated in the cultures. Pathologic examination of the brain tissue revealed necrosis of neuron cells and broad hyperplasia of gliocytes. Immunostaining demonstrated that monokine induced by interferon-Γ (Mig) was expressed in gliocytes with the infiltration of CD68+ monocytes/macrophages and CD3+ T lymphocytes in the brain mesenchyme. Cytokine/chemokine assay revealed that levels of interferon-Γ–inducible protein 10 and Mig in the blood were highly elevated, although the levels of other cytokines and chemokines were close to normal. Conclusions. This study provides direct evidence that SARS human coronavirus is capable of infecting the central nervous system, and that Mig might be involved in the brain immunopathology of SARS.
Angelman syndrome (AS) is a rare neurodevelopmental disorder characterized by developmental delay, impaired communication, motor deficits and ataxia, intellectual disabilities, microcephaly, and ...seizures. The genetic cause of AS is the loss of expression of UBE3A (ubiquitin protein ligase E6-AP) in the brain, typically due to a deletion of the maternal 15q11-q13 region. Previous studies have been performed using a mouse model with a deletion of a single exon of Ube3a. Since three splice variants of Ube3a exist, this has led to a lack of consistent reports and the theory that perhaps not all mouse studies were assessing the effects of an absence of all functional UBE3A. Herein, we report the generation and functional characterization of a novel model of Angelman syndrome by deleting the entire Ube3a gene in the rat. We validated that this resulted in the first comprehensive gene deletion rodent model. Ultrasonic vocalizations from newborn Ube3a
were reduced in the maternal inherited deletion group with no observable change in the Ube3a
paternal transmission cohort. We also discovered Ube3a
exhibited delayed reflex development, motor deficits in rearing and fine motor skills, aberrant social communication, and impaired touchscreen learning and memory in young adults. These behavioral deficits were large in effect size and easily apparent in the larger rodent species. Low social communication was detected using a playback task that is unique to rats. Structural imaging illustrated decreased brain volume in Ube3a
and a variety of intriguing neuroanatomical phenotypes while Ube3a
did not exhibit altered neuroanatomy. Our report identifies, for the first time, unique AS relevant functional phenotypes and anatomical markers as preclinical outcomes to test various strategies for gene and molecular therapies in AS.
Summary
What is Known and Objective: The pathogenic mechanism of antituberculosis drug‐induced hepatotoxicity (ATDH) is thought to involve drug‐metabolizing enzymes including N‐acetyl transferase2 ...(NAT2), cytochrome P4502E1 (CYP2E1) and glutathione S‐transferase (GST) M1, T1. The associations between genetic polymorphisms of those genes and ATDH have been reported but with inconsistent results. Moreover, most studies were hospital‐based retrospective studies and not prospective. We aimed to investigate possible associations of CYP2E1, GSTM1 and GSTT1 genetic polymorphisms with ATDH using a more robust case–control study nested in a population‐based prospective antituberculosis treatment cohort.
Methods: A total of 4304 patients with smear‐positive tuberculosis (TB) who received standard short‐course chemotherapy were monitored for 6–9 months. Incidence density sampling method was adopted to select controls and 4 : 1 matched with each ATDH cases by age (±5 years), sex, treatment history, disease severity and drug dosage. The CYP2E1, GSTM1 and GSTT1 polymorphisms were genotyped using PCR–RFLP and multiplex PCR methods. Conditional logistic regression model was used to calculate odds ratio (OR) and 95% confidence interval (CI), as well as corresponding P‐values.
Results and Discussion: A total of 89 ATDH cases and 356 controls were included in this study. There was no statistically significant association between CYP2E1 RsaI c1/c1 genotype or DraI C/C genotype and ATDH (OR = 0·99, 95% CI:0·62–1·59; OR = 1·13, 95% CI: 0·40–3·20, respectively) compared with CYP2E1 RsaI c1/c2 or c2/c2 genotypes or DraI D/D genotype, or between GSTM1/GSTT1 null genotypes and ATDH (OR = 1·22, 95% CI: 0·76–1·96; OR = 0·96, 95% CI: 0·60–1·52, respectively) compared with non‐null genotypes.
What is new and Conclusion: This is the first study of the involvement of CYP2E1, GSTM1 and GSTT1 genetic polymorphisms in ATDH using a nested case–control population‐based prospective cohort design. We could not confirm positive associations of genetic polymorphisms of CYP2E1 RsaI, CYP2E1 DraI, GSTM1 null and GSTT1 null with ATDH reported by various groups, in our Chinese TB population.
A heterodimer of ultraspiracle (USP) and ecdysone receptor (EcR) mediates 20‐hydroxyecdysone (20E) signalling cascade to regulate insect moulting and metamorphosis. However, at least two questions ...remain to be addressed in terms of the molecular importance of USP in insect species. First, is USP involved in both regulation of ecdysteroidogenesis and mediation of 20E signalling in non‐drosophilid insects, as in Drosophila melanogaster? Second, does USP play any role in larval metamorphosis except as the partner of heterodimeric receptor to activate the downstream 20E signalling genes? In this paper, we found that RNA interference (RNAi) of LdUSP in the final (fourth) instar larvae reduced the messenger RNA levels of four ecdysteroidogenesis genes (Ldspo, Ldphm, Lddib and Ldsad) and 20E titre, and repressed the expression of five 20E signal genes (EcRA, HR3, HR4, E74 and E75) in Leptinotarsa decemlineata. The LdUSP RNAi larvae remained as prepupae, with developing antennae, legs and discs of forewings and hindwings. Dietary supplement with 20E restored the expression of the five 20E signal genes, but only partially alleviated the decreased pupation rate in LdUSP RNAi beetles. Knockdown of LdUSP at the penultimate (third) instar larvae did not affect third–fourth instar moulting. However, silencing LdUSP caused similar but less severe impairments on pupation. Accordingly, we propose that USP is undoubtedly necessary for ecdysteroidogenesis, for mediation of 20E signalling and for initiation of metamorphosis in L. decemlineata.
In order to better understand the Mesozoic tectonic evolution of Southeast China Block (SECB in short), this paper describes geological features of Mesozoic basins that are widely distributed in the ...SECB. The analyzed data are derived from a regional geological investigation on various Mesozoic basins and a recently compiled 1:1,500,000 geological map of Mesozoic–Cenozoic basins. Two types of basin are distinguished according to their tectonic settings, namely, the post-orogenic basin (Type I) and the intracontinental extensional basin (Type II); the latter includes the graben and the half-graben or faulted-depression basins. Our studies suggest that the formation of these basins connects with the evolution of geotectonics of the SECB. The post-orogenic basin (Type I) was formed in areas from the piedmont to the intraland during the interval from Late Triassic to Early Jurassic; and the formation of the intracontinental extensional basin (Type II) connects with an intracontinental crustal thinning setting in the Late Mesozoic. The graben basin was generated during the Middle Jurassic and is associated with a bimodal volcanic eruption; and the half-graben or faulted-depression basin, filled mainly by the rhyolite, tuff and sedimentary rocks during Early Cretaceous, is occupied by the Late Cretaceous–Paleogene red-colored terrestrial clastic rocks. We noticed that the modern outcrops of numerous granites and basins occur in a similar level, and the Mesozoic granitic bodies contact with the adjacent basins by large normal faults, suggesting that the modern landforms between granites and basins were yielded by the late crustal movement. The modern basin and range framework was settled down in the Cretaceous. Abundant sedimentary structures are found in the various basins, from that the deposited environments and paleo-currents are concluded; during the Late Triassic–Early Jurassic time, the source areas were situated to the north and northeast sides of the outcrop region. In this paper, we present the study results on one geological and geographical separating unit and two separating fault zones. The Wuyi orogenic belt is a Late Mesozoic paleo-geographically separating unit, the Ganjiang fault zone behaves as the western boundary of Early Cretaceous volcanic rocks, and the Zhenghe–Dapu fault zone separates the SE-China Coastal Late Mesozoic volcanic-sedimentary basins and the Wuyi orogenic belt. Finally, we discuss the geodynamic mechanisms forming various basins, proposing a three-stage model of the Mesozoic sedimentary evolution.
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