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Tumor-derived exosomes and their contents promote cancer metastasis. Phosphoglycerate mutase 1 (PGAM1) is involved in various cancer-related processes. Nevertheless, the underlying mechanism of exosomal PGAM1 in prostate cancer (PCa) metastasis remains unclear. In this study, we performed in vitro and in vivo to determine the functions of exosomal PGAM1 in the angiogenesis of patients with metastatic PCa. We performed Glutathione-S-transferase pulldown, co-immunoprecipitation, western blotting and gelatin degradation assays to determine the pathway mediating the effect of exosomal PGAM1 in PCa. Our results revealed a significant increase in exosomal PGAM1 levels in the plasma of patients with metastatic PCa compared to patients with non-metastatic PCa. Furthermore, PGAM1 was a key factor initiating PCa cell metastasis by promoting invadopodia formation and could be conveyed by exosomes from PCa cells to human umbilical vein endothelial cells (HUVECs). In addition, exosomal PGAM1 could bind to γ-actin (ACTG1), which promotes podosome formation and neovascular sprouting in HUVECs. In vivo results revealed exosomal PGAM1 enhanced lung metastasis in nude mice injected with PCa cells via the tail vein. In summary, exosomal PGAM1 promotes angiogenesis and could be used as a liquid biopsy marker for PCa metastasis.
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Exossomos , MicroRNAs , Neoplasias da Próstata , Animais , Humanos , Masculino , Camundongos , Actinas/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Células Endoteliais/metabolismo , Exossomos/metabolismo , Camundongos Nus , MicroRNAs/metabolismo , Metástase Neoplásica/patologia , Fosfoglicerato Mutase/genética , Fosfoglicerato Mutase/metabolismo , Neoplasias da Próstata/patologiaRESUMO
Introduction: It is well known that the common chimpanzee, as both the closest living relative to humans and a facultative bipedal, has the capability of bipedal standing but cannot do so fully upright. Accordingly, they have been of exceeding significance in elucidating the evolution of human bipedalism. There are many reasons why the common chimpanzee can only stand with its hips-knees bent, such as the distally oriented long ischial tubercle and the almost absent lumbar lordosis. However, it is unknown how the relative positions of their shoulder-hip-knee-ankle joints are coordinated. Similarly, the distribution of the biomechanical characteristics of the lower-limb muscles and the factors that affect the erectness of standing as well as the muscle fatigue of the lower limbs remain a mystery. The answers are bound to light up the evolutional mechanism of hominin bipedality, but these conundrums have not been shed much light upon, because few studies have comprehensively explored the effects of skeletal architecture and muscle properties on bipedal standing in common chimpanzees. Methods: Thus, we first built a musculoskeletal model comprising the head-arms-trunk (HAT), thighs, shanks, and feet segments of the common chimpanzee, and then, the mechanical relationships of the Hill-type muscle-tendon units (MTUs) in bipedal standing were deduced. Thereafter, the equilibrium constraints were established, and a constrained optimization problem was formulated where the optimization objective was defined. Finally, thousands of simulations of bipedal standing experiments were performed to determine the optimal posture and its corresponding MTU parameters including muscle lengths, muscle activation, and muscle forces. Moreover, to quantify the relationship between each pair of the parameters from all the experimental simulation outcomes, the Pearson correlation analysis was employed. Results: Our results demonstrate that in the pursuit of the optimal bipedal standing posture, the common chimpanzee cannot simultaneously achieve maximum erectness and minimum muscle fatigue of the lower limbs. For uni-articular MTUs, the relationship between muscle activation, relative muscle lengths, together with relative muscle forces, and the corresponding joint angle is generally negatively correlated for extensors and positively correlated for flexors. For bi-articular MTUs, the relationship between muscle activation, coupled with relative muscle forces, and the corresponding joint angles does not show the same pattern as in the uni-articular MTUs. Discussion: The results of this study bridge the gap between skeletal architecture, along with muscle properties, and biomechanical performance of the common chimpanzee during bipedal standing, which enhances existing biomechanical theories and advances the comprehension of bipedal evolution in humans.
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PURPOSEï¼The purpose of this study was to evaluate the effect of various storage methods on shear bond strength of enamel of bovine teeth and find the storage condition that could preserve the similar bond strength as the freshly extracted teeth. METHODS: One hundred and thirty freshly extracted bovine teeth were divided into 13 groups. One was the reference group and 12 were the experimental group. Each group contained 10 teeth. Teeth in the reference group were operated on the same day as the teeth were extracted, while teeth in the experimental groups were stored in different methods (4% formaldehyde solution at 4 â, 23 â, 1% chloramine T at 4 â, 23 â, distilled water at 4 â, 23 â). After stored for 30 days and 90 days, the bovine teeth were taken out and then the shear bond strength was tested. The data were analyzed with SPSS 20.0 software package. RESULTS: The bovine teeth stored in 4% formaldehyde and 1% chloramine T at 23 â and in distilled water at 4 â achieved similar bond strength as freshly extracted teeth at 30 days and 90 days, and the bond strength did not change over time. The bovine teeth stored in 4% formaldehyde solution and 1% chloramine T at 4 â at 30 days had higher shear bond strength than freshly extracted bovine teeth, but over time the bond strength reduced and reached the similar level at 90 days. The bovine teeth stored in distilled water at 23 â obtained similar bond strength as freshly extracted teeth at 30 days but over time the bond strength reduced until 90 days. CONCLUSIONS: Bovine teeth stored in 4% formaldehyde solution and 1% chloramine T at 23 â and in distilled water at 4 â achieved similar bond strength as freshly extracted teeth and does not change over time. These three methods are recommended for storing bovine teeth.
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Colagem Dentária , Bovinos , Animais , Colagem Dentária/métodos , Esmalte Dentário , Formaldeído/química , Água/química , Resistência ao Cisalhamento , Cimentos de Resina , Teste de Materiais , Análise do Estresse DentárioRESUMO
Somatic cell nuclear transfer (SCNT) in pigs is a promising technology in biomedical research by association with transgenesis for xenotransplantation and disease modeling technologies. Handmade cloning (HMC) is a simplified SCNT method that does not require micromanipulators and facilitates the generation of cloned embryos in large quantities. As a result of HMC fine-tuning for porcine-specific requirements of both oocytes and embryos, HMC has become uniquely efficient (>40% blastocyst rate, 80-90% pregnancy rates, 6-7 healthy offspring per farrowing, and with negligible losses and malformations). Therefore, this chapter describes our HMC protocol to obtain cloned pigs.
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Clonagem de Organismos , Técnicas de Transferência Nuclear , Gravidez , Feminino , Suínos , Animais , Clonagem de Organismos/métodos , Oócitos , Blastocisto , Clonagem MolecularRESUMO
The self-starting performance of a figure-9 fiber laser is critically dependent on the phase shift difference between the counter-propagating beams. Herein, we propose an effective approach to dynamically control the phase shift difference in a figure-9 fiber laser by utilizing the thermal nonlinearity of graphene-decorated microfiber device. With the adjustment of the control laser power injected into the graphene-decorated microfiber, the self-starting mode-locked threshold of the figure-9 fiber laser can be attained in a flexible pump power range, i.e., from 300â mW to 390â mW. These findings demonstrated that the graphene-decorated microfiber could act as a dynamical control device of phase shift difference for improving the performance of figure-9 fiber lasers, and might also open up new possibilities for applications of microfiber photonic devices in the field of ultrafast optics.
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Thunberg fritillary (Fritillaria thunbergii), a perennial used in traditional Chinese herbal medicine, is a members of the family Liliaceae. The degeneration of germplasm is a severe problem in the production of Fritillaria thunbergii var. chekiangensis. However, no information about viral infections of F. thunbergii var. chekiangensis has been reported. In this study, we sequenced the small RNAs of F. thunbergii var. chekiangensis from leaves and bulbs, and viruses were identified using a phylogenetic analysis and BLAST search for sequence. In addition, multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) was used to rapidly detect viruses in this variety. Our study first reported that five viruses infected F. thunbergii var. chekiangensis. Among them, fritillary virus Y (FVY), lily mottle virus (LMoV), Thunberg fritillary mosaic virus (TFMV), and hop yellow virus (HYV) had been reported in F. thunbergii, while apple stem grooving virus was first reported in the genus Fritillaria. A multiplex RT-PCR method was developed to rapidly test the four viruses FVY, LMoV, TFMV, and HYV in F. thunbergii var. chekiangensis. Our results provide a better understanding of the infection of F. thunbergii var. chekiangensis by viruses and a basic reference for the better design of suitable control measures.
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The optimum femtosecond laser direct writing of Bragg gratings on silica optical waveguides has been investigated. The silica waveguide has a 6.5 × 6.5 µm2 cross-sectional profile with a 20-µm-thick silicon dioxide cladding layer. Compared with conventional grating inscribed on fiber platforms, the silica planar waveguide circuit can realize a stable performance as well as a high-efficiency coupling with the fiber. A thin waveguide cladding layer also facilitates laser focusing with an improved spherical aberration. Different from the circular fiber core matching with the Gaussian beam profile, a 1030-nm, 400-fs, and 190-nJ laser is optimized to focus on the top surface of the square silica waveguide, and the 3rd-order Bragg gratings are inscribed successfully. A 1.5-mm long uniform Bragg gratings structure with a reflectivity of 90% at a 1548.36-nm wavelength can be obtained. Cascaded Bragg gratings with different periods are also inscribed in the planar waveguide. Different reflection wavelengths can be realized, which shows great potential for wavelength multiplexing-related applications such as optical communications or sensing.
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OBJECTIVE: Chronic nonbacterial prostatitis (CNP) has remained one of the most prevalent urological diseases, particularly in older men. Dihydroartemisinin (DHA) has been identified as a semi-synthetic derivative of artemisinin that exhibits broad protective effects. However, the role of DHA in inhibiting CNP inflammation and prostatic epithelial cell proliferation remains largely unknown. MATERIALS AND METHODS: CNP animal model was induced by carrageenan in C57BL/6 mouse. Enzyme linked immunosorbent assay (ELISA), Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to examine inflammatory cytokines and proliferation genes expression. Immunofluorescence and immunochemistry staining were used to detect and E2F7 expression. Human prostatic epithelial cells (HPECs) and RWPE-1 was induced by lipopolysaccharide (LPS) to mimic CNP model in vitro. Cell proliferation was determined using MTS assay. RESULTS: DHA significantly alleviated the rough epithelium and inhibited multilamellar cell formation in the prostatic gland cavity and prostatic index induced by carrageenan. In addition, DHA decreased the expression of TNF-α and IL-6 inflammatory factors in prostatitis tissues and in LPS-induced epithelial cells. Upregulation of transcription factor E2F7, which expression was inhibited by DHA, was found in CNP tissues, human BPH tissues and LPS-induced epithelial cells inflammatory response. Mechanically, we found that depletion of E2F7 by shRNA inhibited epithelial cell proliferation and LPS-induced inflammation while DHA further enhance these effects. Furthermore, HIF1α was transcriptional regulated by E2F7 and involved in E2F7-inhibited CNP and cellular inflammatory response. Interestingly, we found that inhibition of HIF1α blocks E2F7-induced cell inflammatory response but does not obstruct E2F7-promoted cell growth. CONCLUSION: The results revealed that DHA inhibits the CNP and inflammation by blocking the E2F7/HIF1α pathway. Our findings provide new evidence for the mechanism of DHA and its key role in CNP, which may provide an alternative solution for the prevention and treatment of CNP.
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Prostatite , Idoso , Animais , Artemisininas , Carragenina/efeitos adversos , Fator de Transcrição E2F7 , Células Epiteliais/metabolismo , Humanos , Inflamação/metabolismo , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Prostatite/induzido quimicamente , Prostatite/tratamento farmacológico , Prostatite/genéticaRESUMO
Chemoresistance and migration represent major obstacles in the therapy of non-small-cell lung cancer (NSCLC), which accounts for approximately 85% of lung cancer patients in clinic. In the present study, we report that the compound C1632 is preferentially distributed in the lung after oral administration in vivo with high bioavailability and limited inhibitory effects on CYP450 isoenzymes. We found that C1632 could simultaneously inhibit the expression of LIN28 and block FGFR1 signalling transduction in NSCLC A549 and A549R cells, resulting in significant decreases in the phosphorylation of focal adhesion kinase and the expression of matrix metalloproteinase-9. Consequently, C1632 effectively inhibited the migration and invasion of A549 and A549R cells. Meanwhile, C1632 significantly suppressed the cell viability and the colony formation of A549 and A549R cells by inhibiting DNA replication and inducing G0/G1 cell cycle arrest. Interestingly, compared with A549 cells, C1632 possesses the same or even better anti-migration and anti-proliferation effects on A549R cells, regardless of drug resistance. In addition, C1632 also displayed the capacity to inhibit the growth of A549R xenograft tumours in mice. Altogether, these findings reveal the potential of C1632 as a promising anti-NSCLC agent, especially for chemotherapy-resistant NSCLC treatment.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Células A549 , Animais , Apoptose , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Camundongos , Proteínas de Ligação a RNA/metabolismo , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismo , Transdução de SinaisRESUMO
How to generate anthropomorphic reaching movement remains a challenging problem in service robots and human motor function repair/reconstruction equipment. However, there is no universally accepted computational model in the literature for reproducing the motion of the human upper limb. In response to the problem, this article presents a computational framework for generating reaching movement endowed with human motion characteristics that imitated the mechanism in the control and realization of human upper limb motions. This article first establishes the experimental paradigm of human upper limb functional movements and proposes the characterization of human upper limb movement characteristics and feature movement clustering methods in the joint space. Then, according to the specific task requirements of the upper limb, combined with the human sensorimotor model, the estimation method of the human upper limb natural postures was established. Next, a continuous task parametric model matching the characteristic motion class is established by using the Gaussian mixture regression method. The anthropomorphic motion generation method with the characteristics of the smooth trajectory and the ability of natural obstacle avoidance is proposed. Finally, the anthropomorphic motion generation method proposed in this article is verified by a human-like robot. The measurement index of the human-likeness degree of the trajectory is given. The experimental results show that for all four tested tasks, the human-likeness degrees were greater than 90.8%, and the trajectories' jerk generated by this method is very similar to the trajectories' jerk of humans, which validates the proposed method.
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Robótica , Humanos , Robótica/métodos , Extremidade Superior/fisiologia , Movimento/fisiologia , PosturaRESUMO
OBJECTIVE: To investigate the expression of phosphoglycerate mutase 1 (PGAM1) in the mouse testis after exposure to single heat stress (SHS). METHODS: We randomly assigned 32 C57 male mice to an SHS (n = 16) and a control group (n = 16), the former bathed in water at 43 â and the latter at 25 â for 15 minutes. At 1 and 7 days after exposure, we harvested the testicular tissue for observation of the morphological changes of testicular cells by HE staining and determination of the location and expression of the PGAM1 protein by immunohistochemistry and Western blot. RESULTS: The testis volume of the mice were reduced significantly, the spermatogenic tubules were disorganized, and the cells were reduced in number after heat stress and basically disappeared after 7 days. Immunohistochemistry showed extensive expression of the PGAM1 protein in the testicular spermatogenic tubules of the SHS-exposed mice, significantly higher than in the control group at 1 day after exposure, which was down-regulated in the testis tissue at 7 days, but still markedly higher than that in the control. Western blot exhibited significantly up-regulated expression of the PGAM1 protein after heat stress compared with that in the control group. CONCLUSIONS: The expression of the PGAM1 protein undergoes dynamic changes in the mouse testis after exposed to single heat stress, which is related to heat stress-induced proliferation and division of testicular spermatogenic cells.
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Fosfoglicerato Mutase , Testículo , Animais , Resposta ao Choque Térmico , Masculino , CamundongosRESUMO
Three mononuclear, dinuclear and one-dimensional dysprosium(III) complexes based on 3-azotriazolyl-2,6-dihydroxybenzoic acid (H4ATB) of [Dy(H3ATB)3]·3H2O (1), [Dy2(H2ATB)2(H2DHB)2(H2O)4]·2CH3CN·5H2O (2), and [Dy2(H2ATB)2(DCB)(DMF)2(H2O)2]·4DMF (3) were synthesized and structurally characterized by X-ray single crystal diffraction (H3DHB = 2,6-dihydroxybenzoic acid, H2DCB = 1,4-dicarboxybenzene). Complex 1 was used as a precursor to synthesize complexes 2 and 3, and 2 was further used to synthesize 3. Complex 1 is a mononuclear complex, in which the Dy(III) ion is in a nine-coordinated structure surrounded by three tridentate chelate H3ATB- ligands. Complex 2 displays a dinuclear structure bridged by two µ2 carboxyl groups of two H2DHB- ligands and two µ1,1-O atoms from the phenolic hydroxyl groups of two H2ATB2- ligands. Complex 3 shows a one-dimensional structure formed by two bridging DCB2- ligands. The magnetic measurements were performed on three complexes 1-3, and they showed different magnetic behavior. Complex 1 shows a field-induced slow magnetic relaxation. Complexes 2 and 3 display distinct slow magnetic relaxation under zero dc field with energy barriers (Ueff) of 26(2) cm-1 and 11(1) cm-1, respectively. The magnetic behavior of three complexes 1-3 was investigated by ab initio calculations.
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The aim of this brief report is to offer a solution for a problem that compromises the quality of in vitro-produced mammalian embryos. The harmful effects of evaporation-induced osmotic changes in mammalian embryo cultures have been recognized only recently. In this technical report, we describe a modified embryo culture dish (Humdish) that provides consistent >97% humidity and fully eliminates osmotic changes in the commonly used drop-under-oil culture systems from day 0 to 6. As an additional benefit, the Humdish also increases the temperature stability of cultures. If subsequent laboratory and clinical experiments prove its value, our suggested approach may help to improve the in vitro environment and quality of all preimplantation stage mammalian embryos, including the most sensitive ones produced from artificial gametes or by somatic cell nuclear transfer.
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Meios de Cultura/normas , Técnicas de Cultura Embrionária/métodos , Técnicas de Cultura Embrionária/normas , Embrião de Mamíferos/citologia , Umidade , Animais , Humanos , Concentração OsmolarRESUMO
Two ion-pair Fe(iii) complexes (PPh4)[FeIII(HATD)2]·2H2O (1, H3ATD = azotetrazolyl-2,7-dihydroxynaphthalene) and [FeII(phen)3][FeIII(HATD)2]2·3DMA·3.5H2O (2, phen = 1,10-phenanthroline, DMA = N,N-dimethylformamide) were synthesized by employing the tridentate ligand H3ATD. Crystal structure analyses reveal that complexes 1 and 2 consist of FeIII ions in an octahedral environment where a FeIII ion is coordinated by two HATD2- ligands forming the [FeIII(HATD)2]- core. The shortest cationanion distance between the phosphorus ion of the (PPh4)+ cation and the ferric ion of the [FeIII(HATD)2]- anion is 13.190 Å in complex 1, whereas that between the ferrous ion of the [FeII(Phen)3]2+ cation and the ferric ion of the [FeIII(HATD)2]- anion is 7.821 Å in complex 2. C-HC and C-HO hydrogen interactions between the [FeII(phen)3]2+ cation and the [FeIII(HATD)2]- anion are observed in 2. Face-to-face π-π stacking interactions between naphthalene rings with the separated interplanar center to center distances of 3.421-3.680 Å were observed, which result in a one-dimensional supramolecular chain in complexes 1 and 2. Magnetic measurements show that complex 1 is in the low-spin (LS) state below 500 K, whereas 2 undergoes a high temperature spin crossover (SCO) between 360 and 500 K. Magneto-structural relationship studies reveal that π-stacking, hydrogen interactions and Coulomb interactions between the [FeIII(HATD)2]- anion and the [FeII(phen)3]2+ cation play a crucial role in the high temperature Fe(iii) SCO behaviour of complex 2.
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Although in vitro culture of human embryos is a crucial step in assisted reproduction, the lack of focused research hampers worldwide standardisation and consistent outcomes. Only 1.2% of research papers published in five leading journals in human reproduction in 2019 focused on in vitro culture conditions, creating the impression that the optimisation process has approached its limits. On the other hand, in vitro culture of mammalian embryos is based on old principles, while there is no consensus on basic issues as density, time, medium change, gas atmosphere and small technical details including the way of drop preparation. This opinion paper aims to highlight and analyse the slow advancement in this field and stimulate research for simple and affordable solutions to meet the current requirements. A possible way for advancement is discussed in detail. Selection of embryos with the highest developmental competence requires individual culture and modification of the widely used "drop under oil" approach. Current use of three-dimensional surfaces instead of large flat bottoms is restricted to time-lapse systems, but these wells are designed for optical clarity, not for the needs of embryos. The size and shape of the original microwells (Well of the Well; WOW) offer a practical and straightforward solution to combine the benefits of communal and individual incubation and improve the overall quality of cultured embryos.
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Blastocisto/citologia , Técnicas de Cultura Embrionária/métodos , Técnicas de Cultura Embrionária/normas , Embrião de Mamíferos/citologia , Desenvolvimento Embrionário , Fertilização in vitro/métodos , Meios de Cultura , HumanosRESUMO
The interaction between LncRNA and RNA-binding protein (RBPs) plays an essential role in the regulation over the malignant progression of tumors. Previous studies on the mechanism of SNHG1, an emerging lncRNA, have primarily focused on the competing endogenous RNA (ceRNA) mechanism. Nevertheless, the underlying mechanism between SNHG1 and RBPs in tumors remains to be explored, especially in prostate cancer (PCa). SNHG1 expression profiles in PCa were determined through the analysis of TCGA data and tissue microarray at the RNA level. Gain- and loss-of-function experiments were performed to investigate the biological role of SNHG1 in PCa initiation and progression. RNA-seq, immunoblotting, RNA pull-down and RNA immunoprecipitation analyses were utilized to clarify potential pathways with which SNHG1 might be involved. Finally, rescue experiments were carried out to further confirm this mechanism. We found that SNHG1 was dominantly expressed in the nuclei of PCa cells and significantly upregulated in PCa patients. The higher expression level of SNHG1 was dramatically correlated with tumor metastasis and patient survival. Functionally, overexpression of SNHG1 in PCa cells induced epithelial-mesenchymal transition (EMT), accompanied by down-regulation of the epithelial marker, E-cadherin, and up-regulation of the mesenchymal marker, vimentin. Increased proliferation and migration, as well as accelerated xenograft tumor growth, were observed in SNHG1-overexpressing PCa cells, while opposite effects were achieved in SNHG1-silenced cells. Mechanistically, SNHG1 competitively interacted with hnRNPL to impair the translation of protein E-cadherin, thus activating the effect of SNHG1 on the EMT pathway, eventually promoting the metastasis of PCa. Our findings demonstrate that SNHG1 is a positive regulator of EMT activation through the SNHG1-hnRNPL-CDH1 axis. SNHG1 may serve as a novel potential therapeutic target for PCa.
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Antígenos CD/metabolismo , Caderinas/metabolismo , Regulação Neoplásica da Expressão Gênica/genética , Neoplasias da Próstata/genética , RNA Longo não Codificante/metabolismo , Humanos , Masculino , Metástase Neoplásica , Neoplasias da Próstata/patologiaRESUMO
A series of innovative thermosetting polymer nanocomposites comprising of polysiloxane-imide-containing benzoxazine (PSiBZ) as the matrix and double-decker silsesquioxane (DDSQ) epoxy or polyhedral oligomeric silsesquioxane (POSS) epoxy were prepared for improving thermosetting performance. Thermomechanical and dynamic mechanical characterizations indicated that both DDSQ and POSS could effectively lower the coefficient of thermal expansion by up to approximately 34% and considerably increase the storage modulus (up to 183%). Therefore, DDSQ and POSS are promising materials for low-stress encapsulation for electronic packaging applications.
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A parasitoid's decision to reject or accept a potential host is fundamental to its fitness. Superparasitism, in which more than one egg of a given parasitoid species can deposit in a single host, is usually considered sub-optimal in systems where the host is able to support the development of only a single parasitoid. It follows that selection pressure may drive the capacity for parasitoids to recognize parasitized hosts, especially if there is a fitness cost of superparasitism. Here, we used microsatellite studies of two distinct populations of Cotesia vestalis to demonstrate that an egg laid into a diamondback moth (Plutella xylostella) larva that was parasitized by a conspecific parasitoid 10 min, 2 or 6 h previously was as likely to develop and emerge successfully as was the first-laid egg. Consistent with this, a naive parasitoid encountering its first host was equally likely to accept a healthy larva as one parasitized 10 min prior, though handling time of parasitized hosts was extended. For second and third host encounters, parasitized hosts were less readily accepted than healthy larvae. If 12 h elapsed between parasitism events, the second-laid egg was much less likely to develop. Discrimination between parasitized and healthy hosts was evident when females were allowed physical contact with hosts, and healthy hosts were rendered less acceptable by manual injection of parasitoid venom into their hemolymph. Collectively, these results show a limited capacity to discriminate parasitized from healthy larvae despite a viability cost associated with failing to avoid superparasitism.