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1.
Artigo em Inglês | MEDLINE | ID: mdl-32114660

RESUMO

Seasonal estrus is a key factor limiting animal fertility, and understanding the molecular mechanisms that regulate animal estrus is important for improving animal fertility. The pituitary gland, which is the most important endocrine gland in mammals, plays an important role in regulating the physiological processes such as growth, development, and reproduction of animals. Here, we used RNA-seq technology to study the expression profile of lncRNAs in the anterior pituitary of sheep during estrus and anestrus. In this study, we identified a total of 995 lncRNAs, of which 335 lncRNAs were differentially expressed in two states (including 38 up-regulated and 297 down-regulated lncRNAs). RT-qPCR verified the expression levels of several lncRNAs. Target predictive analysis revealed that these lncRNAs can act in cis or trans and regulate the expression of genes involved in the regulation of sheep estrus. Target gene enrichment analysis of differentially expressed lncRNAs indicates that these lncRNAs can regulate sheep estrus by regulating hormone metabolism and energy metabolism. Through our research, we provide the expression profile of lncRNAs in the pituitary of sheep, which provides a valuable resource for further understanding of the genetic regulation of seasonal estrus in sheep from the perspective of lncRNAs.

2.
Appl Environ Microbiol ; 86(7)2020 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-32005731

RESUMO

In Saccharomyces cerevisiae, Y family DNA polymerase Rev1 is involved in the repair of DNA damage by translesion DNA synthesis (TLS). In the current study, to elucidate the role of Rev1 in oxidative stress-induced DNA damage in S. cerevisiae, REV1 was deleted and overexpressed; transcriptome analysis of these mutants along with the wild-type strain was performed to screen potential genes that could be associated with REV1 during response to DNA damage. When the yeast cells were treated with 2 mM H2O2, the deletion of REV1 resulted in a 1.5- and 2.8-fold decrease in the survival rate and mutation frequency, respectively, whereas overexpression of REV1 increased the survival rate and mutation frequency by 1.1- and 2.9-fold, respectively, compared to the survival rate and mutation frequency of the wild-type strain. Transcriptome and phenotypic analyses identified that Sml1 aggravated oxidative stress in the yeast cells by inhibiting the activity of Rev1. This inhibition was due to the physical interaction between the BRCA1 C terminus (BRCT) domain of Rev1 and amino acid residues 36 to 70 of Sml1; the cell survival rate and mutation frequency increased by 1.8- and 3.1-fold, respectively, when this interaction was blocked. We also found that Sml1 inhibited Rev1 phosphorylation under oxidative stress and that deletion of SML1 increased the phosphorylation of Rev1 by 46%, whereas overexpression of SML1 reduced phosphorylation of Rev1. Overall, these findings demonstrate that Sml1 could be a novel regulator that mediates Rev1 dephosphorylation to inhibit its activity during oxidative stress.IMPORTANCE Rev1 was critical for cell growth in S. cerevisiae, and the deletion of REV1 caused a severe growth defect in cells exposed to oxidative stress (2 mM H2O2). Furthermore, we found that Sml1 physically interacted with Rev1 and inhibited Rev1 phosphorylation, thereby inhibiting Rev1 DNA antioxidant activity. These findings indicate that Sml1 could be a novel regulator for Rev1 in response to DNA damage by oxidative stress.

3.
Cell Death Dis ; 11(2): 140, 2020 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-32080168

RESUMO

Cardiac hypertrophy (CH) is an independent risk factor for many cardiovascular diseases, and is one of the primary causes of morbidity and mortality in elderly people. Pathological CH involves excessive protein synthesis, increased cardiomyocyte size, and ultimately the development of heart failure. Myotubularin-related protein 14 (MTMR14) is a member of the myotubularin (MTM)-related protein family, which is involved in apoptosis, aging, inflammation, and autophagy. However, its exact function in CH is still unclear. Herein, we investigated the roles of MTMR14 in CH. We show that MTMR14 expression was increased in hypertrophic mouse hearts. Mice deficient in heart MTMR14 exhibited an aggravated aortic-banding (AB)-induced CH phenotype. In contrast, MTMR14 overexpression prevented pressure overload-induced hypertrophy. At the molecular level, prevention of CH in the absence of MTMR14 involved elevations in Akt pathway components, which are key elements that regulate apoptosis and cell proliferation. These results demonstrate that MTMR14 is a new molecular target for the treatment of CH.

4.
Adv Mater ; : e1907214, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-31999014

RESUMO

Designing highly active and robust electrocatalysts for oxygen evolution reaction (OER) is crucial for many renewable energy storage and conversion devices. Here, self-supported monolithic hybrid electrodes that are composed of bimetallic cobalt-molybdenum nitride nanosheets vertically aligned on 3D and bicontinuous nanoporous gold (NP Au/CoMoNx ) are reported as highly efficient electrocatalysts to boost the sluggish reaction kinetics of water oxidation in alkaline media. By virtue of the constituent CoMoNx nanosheets having large accessible CoMoOx surface with remarkably enhanced electrocatalytic activity and the nanoporous Au skeleton facilitating electron transfer and mass transport, the NP Au/CoMoNx electrode exhibits superior OER electrocatalysis in 1 m KOH, with low onset overpotential (166 mV) and Tafel slope (46 mV dec-1 ). It only takes a low overpotential of 370 mV to reach ultrahigh current density of 1156 mA cm-2 , ≈140-fold higher than free CoMoNx nanosheets. The electrocatalytic performance makes it an attractive candidate as the OER catalyst in the water electrolysis.

5.
Biomaterials ; 230: 119627, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31767445

RESUMO

Cell therapy, the treatment of diseases using living cells, offers a promising clinical approach to treating refractory diseases. The global market for cell therapy is growing rapidly, and there is an increasing demand for automated methods that can produce large quantities of high quality therapeutic cells. Biomaterials can be used during cell production to establish a biomimetic microenvironment that promotes cell adhesion and proliferation while maintaining target cell genotype and phenotype. Here we review recent progress and emerging techniques in biomaterial-assisted cell production. The increasing use of auxiliary biomaterials and automated production methods provides an opportunity to improve quality control and increase production efficiency using standardized GMP-compliant procedures.

6.
J Cell Physiol ; 235(3): 2149-2160, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31389030

RESUMO

miR-222 participates in many cardiovascular diseases, but its effect on cardiac remodeling induced by diabetes is unclear. This study evaluated the functional role of miR-222 in cardiac fibrosis in diabetic mice. Streptozotocin (STZ) was used to establish a type 1 diabetic mouse model. After 10 weeks of STZ injection, mice were intravenously injected with Ad-miR-222 to induce the overexpression of miR-222. miR-222 overexpression reduced cardiac fibrosis and improved cardiac function in diabetic mice. Mechanistically, miR-222 inhibited the endothelium to mesenchymal transition (EndMT) in diabetic mouse hearts. Mouse heart fibroblasts and endothelial cells were isolated and cultured with high glucose (HG). An miR-222 mimic did not affect HG-induced fibroblast activation and function but did suppress the HG-induced EndMT process. The antagonism of miR-222 by antagomir inhibited HG-induced EndMT. miR-222 regulated the promoter region of ß-catenin, thus negatively regulating the Wnt/ß-catenin pathway, which was confirmed by ß-catenin siRNA. Taken together, our results indicated that miR-222 inhibited cardiac fibrosis in diabetic mice via negatively regulating Wnt/ß-catenin-mediated EndMT.

7.
Sci Total Environ ; 706: 136037, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31841842

RESUMO

Spatially continuous satellite data have been widely used to estimate monthly air temperature (Ta). However, it is still not clear whether the estimated monthly Ta is temporally consistent with observed Ta or not. In this study, the accuracies of interannual variations and temporal trends in estimated monthly Ta were systematically analyzed for Mainland China during 2001-2018. The differences in accuracy among five ways to input data into the model were investigated. The Cubist algorithm and ten variables were used to estimate monthly Ta. It was found that inputting data for the same month into the model can generate more accurate results than inputting all data into the model. Using temporal variables (i.e., month and year) can significantly increase the accuracy of estimated Ta. These results can be explained by different relationships between Ta and auxiliary variables that appear at different times. Thus, using temporal variables can help distinguish between different relationships and improve accuracy levels of the estimated Ta. When applying the best method (inputting data for the same month into the model and using the year as a temporal variable), the coefficient of determination (R2) of estimated monthly mean Ta, interannual variations in monthly mean Ta and temporal trends in monthly mean Ta were recorded as 0.997, 0.731 and 0.848, respectively. The root mean squared errors (RMSEs) of estimated monthly mean Ta, interannual variations in monthly mean Ta and temporal trends in monthly mean Ta were recorded as 0.629 °C, 0.593 °C and 0.201 °C/decade, respectively. An accurate, national coverage, 1 km spatial resolution and long time series (2001-2018) monthly mean, maximum and minimum Ta dataset was finally developed. The dataset can be of great use to many fields such as climatology, hydrology and ecology.

8.
Yonsei Med J ; 61(1): 20-29, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31887796

RESUMO

PURPOSE: Curcumin exerts its anti-cancer effects, partly by targeting special microRNAs, in human cancers. MiR-21 is a key oncomir in carcinogenesis of multiple human cancers. Here, we aimed to further explore the mechanistic insight into the link between curcumin and miR-21 on diffuse large B-cell lymphoma (DLBCL). MATERIALS AND METHODS: Quantitative real-time PCR assays were performed to assess the levels of miR-21 and Von Hippel-Lindau (VHL) mRNA. In situ hybridization assay was used for miR-21 expression visualization in lymphoma tissues. Western blot was used for determination of VHL protein, Ki-67, caspase-3, and cleaved caspase-3 levels. Dual-luciferase reporter assay and RNA immunoprecipitation assay were employed to confirm the direct target of miR-21. MTT assay, flow cytometric analysis, and transwell assay were used to evaluate cell proliferation, apoptosis, and migration and invasion capacities, respectively. RESULTS: Curcumin repressed the proliferation, migration, and invasion abilities and promoted apoptosis in SU-DHL-8 cells. Curcumin inhibited miR-21 expression and curcumin exerted its anti-proliferation, anti-migration, anti-invasion, and pro-apoptosis effects by miR-21 in SU-DHL-8 cells. VHL was a direct target of miR-21. Moreover, curcumin exerted its regulatory effects on SU-DHL-8 cells by VHL. CONCLUSION: Curcumin exerted its anti-proliferation, anti-migration, anti-invasion, and pro-apoptosis functions, at least partly, by repressing miR-21 and regulating VHL expression in DLBCL cell line. Our findings provided a possible molecular mechanism of curcumin-mediated anti-cancer effect.


Assuntos
Apoptose/efeitos dos fármacos , Movimento Celular/ética , Curcumina/farmacologia , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/patologia , MicroRNAs/metabolismo , Proteína Supressora de Tumor Von Hippel-Lindau/metabolismo , Apoptose/genética , Sequência de Bases , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Curcumina/uso terapêutico , Regulação Neoplásica da Expressão Gênica , Humanos , Linfoma Difuso de Grandes Células B/tratamento farmacológico , MicroRNAs/genética , Invasividade Neoplásica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
9.
Biosci Rep ; 39(12)2019 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-31830266

RESUMO

OBJECTIVE: MicroRNA-218 (miR-218) critical for preventing the progression of numerous diseases, including diseases of the retinal pigment epithelium (RPE). However, the mechanism by which miR-218 regulates the PRE in humans remains largely unknown. Our study investigated the effects of glucose-induced miR-218 expression on human RPE cells (ARPE-19), as well as its targeted regulatory effect. METHODS: The levels of miR-218 and runt-related transcription factor 2 (RUNX2) expression were investigated by RT-qPCR or Western blot assays. Cell viability and apoptosis were assessed by CCK-8 assays, flow cytometry, and Hoechst staining. A luciferase reporter assay was performed to determine whether Runx2 is a target gene of miR-218. RESULTS: Our results showed that glucose up-regulated miR-218 expression, suppressed proliferation, and induced the apoptosis of ARPE-19 cells. We verified that miR-218 could inhibit the proliferation and facilitate the apoptosis of ARPE-19 cells, while inhibition of miR-218 expression produced the opposite effects. In terms of mechanism, we demonstrated that RUNX2 was a direct target of miR-218. Functional experiments showed that Runx2 served as a miR-218 target to help inhibit the proliferation and induction of apoptosis in ARPE-19 cells. CONCLUSION: Our findings suggest the miR-218/Runx2 axis as a potential target for treating diabetic retinopathy (DR).

10.
Urol Int ; : 1-6, 2019 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-31852007

RESUMO

OBJECTIVE: To investigate and compare the influence of two numerical detrusor contractility parameters, the bladder contractility index (BCI) and the maximum Watts factor (WFmax), on transurethral resection of the prostate (TURP) outcome. METHODS: A retrospective study was conducted on 236 patients who had undergone urodynamic assessment preoperatively and TURP for benign prostatic obstruction. They were evaluated by International Prostate Symptom Score (IPSS) and uroflowmetry preoperatively and 3 months postoperatively. Related criteria were established to determine the overall efficacy of TURP. Logistic regression analysis and receiver operating characteristic curves were made to investigate the influence of the BCI and WFmax on TURP efficacy. RESULTS: Among the 236 patients, 195 treatments were effective and 41 ineffective. Multivariate analysis showed that both the BCI (OR 1.038) and the WFmax (OR 1.291) could influence TURP efficacy. For predicting TURP efficacy, the optimal cut-off values of the BCI and WFmax were 98.7 and 10.27 W/m2, respectively. The AUC, sensitivity and specificity of the BCI were 0.722, 78.5% and 61.0%; those of the WFmax were 0.761, 73.9% and 73.2%, with no significant difference (p > 0.05). CONCLUSIONS: To some extent, the BCI and the WFmax can predict TURP efficacy equally well. A discrimination level of 10.27 W/m2 may be a threshold value for detrusor underactivity (DU); as regards the BCI, the current threshold value is appropriate to diagnose DU.

11.
Int J Biol Macromol ; 2019 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-31739042

RESUMO

The development of skeletal muscle is a complex biological process involving a variety of regulatory pathways. A deeper understanding of the developmental mechanisms of skeletal muscle is of great significance for the treatment of muscular lesions. In recent years, studies have shown that circRNAs, as a new class of post-transcriptional regulators, play an important regulatory role in a variety of biological processes, but their function in skeletal muscle development remains unclear. Here, we used C2C12 myoblasts to study circ-HIPK3, which has significant differences in the developmental stages of skeletal muscle and is highly expressed. We found that the expression level of circ-HIPK3 was significantly up-regulated (p < .05) with the persistence of C2C12 cell differentiation. Combining the results of bioinformatics analysis and dual luciferase reporter experiments, we found that circ-HIPK3 is a sponge of miR-124 and miR-379 that regulate muscle differentiation. Our study shows that circ-HIPK3 can promote the differentiation of C2C12 myoblasts, providing a scientific basis for further research on the development of skeletal muscle at the level of circRNAs.

12.
Int J Biol Sci ; 15(12): 2615-2626, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31754334

RESUMO

Background/Aims: Vascular smooth muscle cell (VSMC) hyperplasia plays important roles in the pathogenesis of many vascular diseases, such as atherosclerosis and restenosis. Many microRNAs (miRs) have recently been reported to regulate the proliferation and migration of VSMC. In the current study, we aimed to explore the function of miR-93 in VSMCs and its molecular mechanism. Methods: First, qRT-PCR and immunofluorescence assays were performed to determine miR-93 expression in rat VSMCs following carotid artery injury in vivo and platelet-derived growth factor-BB (PDGF-BB) stimulation in vitro. Next, the biological role of miR-93 in rat VSMC proliferation and migration was examined in vivo and vitro. EdU incorporation assay and MTT assay for measuring cell proliferation, Transwell cell invasion assay and Cell scratch wound assay for measuring cell migration. Then, the targets of miR-93 were identified. Finally, the expression levels of proteins in the Raf-ERK1/2 pathway were measured by western blot. Results: MiR-93 was upregulated in rat VSMCs following carotid artery injury in vivo. Similar results were observed in ex vivo cultured VSMCs after PDGF-BB treatment. MiR-93 inhibition suppressed neointimal formation after carotid artery injury. Moreover, our results demonstrated that a miR-93 inhibitor suppressed the PDGF-BB induced proliferation and migration of in VSMC. This inhibitor also decreased the expression levels of MMP2 and cyclin D1. Mechanistically, we discovered that mitofusin 2(Mfn2) is a direct target of miR-93. Furthermore, an analysis of the signaling events revealed that miR-93-mediated VSMC proliferation and migration occurred via the Raf-ERK1/2 pathway. Conclusions: Our findings suggest that miR-93 promotes VSMCs proliferation and migration by targeting Mfn2. MiR-93 may be a new target for treating in-stent restenosis.

13.
Front Genet ; 10: 674, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31379930

RESUMO

The number of vertebrae, especially thoracic vertebrae, is an important economic trait that may influence carcass length and meat production in animals. However, the genetic basis of vertebrae number in sheep is still poorly understood. To detect the candidate genes, 400 increased number of thoracic vertebrae (T14L6) and 200 normal (T13L6) Kazakh sheep were collected. We generated and sequenced 60 pools of genomic DNA (each pool prepared by mixing genomic DNA from 10 sheep with the same thoracic traits), with an average depth of coverage of 25.65×. We identified a total of 42,075,402 SNPs and 11 putatively selected genomic regions, including the VRTN gene and the HoxA gene family that regulate vertebral development. The most prominent areas of selective elimination were located in a region of chromosome 7, including VRTN, which regulates spinal development and morphology. Further investigation indicated that the expression level of the VRTN gene during fetal development was significantly higher in sheep with more thoracic vertebrae than in those with a normal number of thoracic vertebrae. A genome-wide comparison between sheep with increased and normal numbers of thoracic vertebrae showed that the VRTN gene is the major selection locus for the number of thoracic vertebrae in sheep and has the potential to be utilized in sheep breeding in the future.

14.
Environ Sci Pollut Res Int ; 26(30): 30808-30825, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31444726

RESUMO

Rapid urbanization significantly changes vegetation coverage and heat distribution, which threatens the sustainable development and the quality of life. As the largest developing city in Central China, Wuhan was chosen as the experimental region. This study investigated the urbanization process of Wuhan from 1989 to 2917 based on Landsat data. Combined with MODIS EVI (Enhanced Vegetation Index) and LST (Land Surface Temperature) data, vegetation disturbance and surface urban heat island (SUHI) caused by urbanization were discussed for 2001-2017. Furthermore, correlation between ∆EVI (urban EVI minus rural EVI) and ∆LST (urban LST minus rural LST) was also conducted. The results were as follows: (1) Wuhan experienced a strong urbanization over the past 29 years, with an increasing urban expansion rate and the altered dominant urban expansion pattern (edge expansion and infilling). After the enhanced vegetation functions and urban increased structures, the urbanization finally caused the fragmented patches and irregular urban shapes. (2) Urbanization had a positive effect on LST but a negative effect on EVI. From 2001 to 2017, the highest increasing rate of ∆LST for the old urban area (OUA) and urbanized area (UA) was both observed in summer daytime (OUA, 0.106 °C/a; UA, 0.207 °C/a). The decreasing rate of ∆EVI reached the highest value in summer (OUA, 0.00697/a; UA, 0.00298/a). (3) There was a strong negative correlation (except spring and winter for OUA) between ∆EVI and ∆LST in daytime, which proved that the activity of vegetation in daytime could relieve LST to a certain extent. This study clarifies the dynamic urbanization process of Wuhan and discusses its impacts on vegetation change and SUHI. Efficiently investigating urbanization process and quantifying its impacts on urban environment are critical for regional ecological conservation.


Assuntos
Plantas , Urbanização , China , Cidades , Monitoramento Ambiental/métodos , Tecnologia de Sensoriamento Remoto , Estações do Ano , Temperatura Ambiente
15.
Artigo em Inglês | MEDLINE | ID: mdl-31331887

RESUMO

Model-free tracking is a widely-accepted approach to track an arbitrary object in a video using a single frame annotation with no further prior knowledge about the object of interest. Extending this problem to track multiple objects is really challenging because: a) the tracker is not aware of the objects' type while trying to distinguish them from background (detection task), and b) The tracker needs to distinguish one object from other potentially similar objects (data association task) to generate stable trajectories. In order to track multiple arbitrary objects, most existing model-free tracking approaches rely on tracking each target individually by updating their appearance model independently. Therefore, in this scenario they often fail to perform well due to confusion between the appearance of similar objects, their sudden appearance changes and occlusion. To tackle this problem, we propose to use both appearance and motion models, and to learn them jointly using graphical models and deep neural networks features. We introduce an indicator variable to predict sudden appearance change and/or occlusion. When these happen, our model does not update the appearance model thus avoiding using the background and/or incorrect object to update the appearance of the object of interest mistakenly, and relies on our motion model to track. Moreover, we consider the correlation among all targets, and seek the joint optimal locations for all targets simultaneously as a graphical model inference problem. We learn the joint parameters for both appearance model and motion model in an online fashion under the framework of LaRank. Experiment results show that our method achieved superior performance compared to the competitive methods.

16.
J Tissue Eng Regen Med ; 13(11): 2101-2120, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31359625

RESUMO

Resolving the origin and development of tumor heterogeneity has proven to be a crucial challenge in cancer research. In vitro tumor models have been widely used for both scientific and clinical research. Currently, tumor models based on 2D cell culture, animal models, and 3D cell-laden constructs are widely used. Heterogeneous tumor models, which consist of more than one cell type and mimic cell-cell as well as cell-matrix interactions, are attracting increasing attention. Heterogeneous tumor models can serve as pathological models to study the microenvironment and tumor development such as tumorigenesis, invasiveness, and malignancy. They also provide disease models for drug screening and personalized therapy. In this review, the current techniques, models, and oncological applications regarding 3D heterogeneous tumor models are summarized and discussed.

17.
Cells ; 8(6)2019 06 19.
Artigo em Inglês | MEDLINE | ID: mdl-31248210

RESUMO

CircRNA is a type of closed circular non-coding RNA formed by reverse splicing and plays an important role in regulating the growth and development of plants and animals. To investigate the function of circ-FoxO3 in mouse myoblast cells' (C2C12) differentiation and proliferation, we used RT-qPCR to detect the expression level of circ-FoxO3 in mouse myoblast cells at different densities and different differentiation stages, and the specific interference fragment was used to inhibit the expression level of circ-FoxO3 in myoblast cells to observe its effect on myoblast cells proliferation and differentiation. We found that the expression level of circ-FoxO3 in myoblast cells increased with the prolongation of myoblast cells differentiation time, and its expression level decreased with the proliferation of myoblast cells. At the same time, we found that the differentiation ability of the cells was significantly increased (p < 0.05), but the cell proliferation was unchanged (p > 0.05) after inhibiting the expression of circ-FoxO3 in myoblast cells. Combining the results of bioinformatics analysis and the dual luciferase reporter experiment, we found that circ-FoxO3 is a sponge of miR-138-5p, which regulates muscle differentiation. Our study shows that circ-FoxO3 can inhibit the differentiation of C2C12 myoblast cells and lay a scientific foundation for further study of skeletal muscle development at circRNA levels.


Assuntos
Diferenciação Celular , Mioblastos/citologia , Mioblastos/metabolismo , /metabolismo , Animais , Sequência de Bases , Diferenciação Celular/genética , Linhagem Celular , Proliferação de Células/genética , Regulação da Expressão Gênica , Camundongos Endogâmicos C57BL , MicroRNAs/genética , MicroRNAs/metabolismo , /genética
18.
Zookeys ; 835: 43-63, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31043849

RESUMO

In this study, two new mitochondrial genomes (mitogenomes) of Mesonemourametafiligera and Mesonemouratritaenia from the family Nemouridae (Insecta: Plecoptera) were sequenced. The Mesonemourametafiligera mitogenome was a 15,739 bp circular DNA molecule, which was smaller than that of M.tritaenia (15,778 bp) due to differences in the size of the A+T-rich region. Results show that gene content, gene arrangement, base composition, and codon usage were highly conserved in two species. Ka/Ks ratios analyses of protein-coding genes revealed that the highest and lowest rates were found in ND6 and COI and that all these genes were evolving under purifying selection. All tRNA genes in nemourid mitogenomes had a typical cloverleaf secondary structure, except for tRNASer(AGN) which appeared to lack the dihydrouridine arm. The multiple alignments of nemourid lrRNA and srRNA genes showed that sequences of three species were highly conserved. All the A+T-rich region included tandem repeats regions and stem-loop structures. The phylogenetic analyses using Bayesian inference (BI) and maximum likelihood methods (ML) generated identical results. Amphinemurinae and Nemourinae were sister-groups and the family Nemouridae was placed as sister to Capniidae and Taeniopterygidae.

19.
Acta Biomater ; 95: 245-257, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31128321

RESUMO

A new three-dimensional (3D) cell printing system was developed and investigated to organize multiple cells/biomaterials with a control precision within 100 µm. This system can be used for the in vitro construction of heterogeneous tissue models. The proposed printing system was achieved by the integration of extrusion printing and alternating viscous and inertial force jetting (AVIFJ) techniques using dual-nozzle switching. In this technique, hydrogels containing high cell densities were extruded using extrusion printing, while droplets containing single cells were precisely manipulated using AVIFJ. The droplets that contained single cells were at the scale of pico-liters and could be accurately positioned at the micron scale. Stable hydrogel structures with adjustable diameters were also printed, with cell viabilities exceeding 90% after printing. A heterogeneous tumor model that contained spheroids and human umbilical vein endothelial cells (HUVECs) was then constructed using the established integrated cell printing system in a stepwise or simultaneous fashion. HUVEC-loaded droplets were observed to locate around the preformed tumor spheroids as designed. Cells and spheroids in the model maintained high cell viability and sustained growth throughout the culture period. The ELISA results of albumin production also proved that the spheroids maintained increased cellular function during the culture. These results demonstrated the feasibility of this integrated 3D printing system for the engineering of in vitro heterogeneous tissue models for future biological and pathological studies. STATEMENT OF SIGNIFICANCE: Addressing the challenge of multi-scale printing in the construction of heterogeneous tissue models, a new 3D cell printing system was developed to organize cells/biomaterials of a control precision within 100 µm. AVIFJ was integrated with extrusion printing, thereby achieving the construction of cell interactions between single cells and spheroids, the manipulation of single cells in a 3D microenvironment with high accuracy, and the real-time on-demand printing. The printed heterogeneous tumor model maintained cell viability, sustained cell growth, and increased cell function during 7 days of culture. We believed that this work would benefit the production of functional artificial tissues, enabling the construction of more biomimetic cell arrangements and microenvironment to support cell functions.

20.
Biochem Biophys Res Commun ; 514(2): 490-496, 2019 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-31056262

RESUMO

Long-non-coding RNA small nucleolar RNA host gene 12(SNHG12) was reported to be highly up-regulated in brain microvascular endothelium after cerebral ischemia. Autophagy has been shown to have protective effects against cerebral ischemic insults. However, molecular mechanisms of SNHG12 in regulating autophagy during cerebral ischemia/reperfusion (I/R) injury remain unclear. Here, we established middle cerebral artery occlusion/reperfusion (MCAO/R) model in mice and adopted oxygen-glucose deprivation and reperfusion (OGD/R) SH-SY5Y cell model to mimic cerebral I/R injury in vitro. Triphenyltetrazolium chloride (TTC) staining was used to measure infarct size. Bederson and Longa score systems were used to evaluate neurological behavioral and defects, respectively. CCK-8, EdU staining, flow cytometry and Hoechst 33258 staining were performed to determine the biological function of SNHG12 on SH-SY5Y cell under OGD/R condition. The autophagy levels were determined by Western blotting and LC3B immunofluorescence. We found the expression of SNHG12 was up-regulated by cerebral I/R in mice andSH-SY5Y cell model after OGD/R. Up-regulated SNHG12 alleviated OGD/R-induced SH-SY5Y cell injury and induced autophagy activation, as indicated by an increased ratio of LC3 II/I and Beclin-1, decreased p62. On the contrary, down-regulation of SNHG12 exacerbated SH-SY5Y cell injury after OGD/R and inhibited autophagy. Furthermore, autophagy activator rapamycin or inhibitor 3-MA partially reversed the down-regulation or up-regulation of SNHG12 effect in OGD/R-inducedSH-SY5Y cell injury, respectively. Taken together, these findings suggest that SNHG12 as an autophagy inducer alleviates cerebral I/R injury, which might be a new therapeutic target of ischemic stroke.

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