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OBJECTIVES: Dense comedones are common in patients with acne vulgaris, and promoting treatment can prevent the progression of acne lesions. However, the efficacy-time conflict makes the treatment challenging and the medication options are limited by the side effects. MATERIALS AND METHODS: Thirty-five patients with symmetrical dense comedones were enrolled and the two sides of the face were randomly assigned to receive 30% supramolecular salicylic acid (SSA) combined with CO2 laser or CO2 laser monotherapy at an interval of 2 weeks for six treatment sessions. Comedones count, porphyrin index (PI), texture index (TI), melanin index, erythema index, hydration index (HI), transepidermal water loss (TEWL), and side effects were recorded at each visit till the 12th week. RESULTS: Thirty-one patients completed the study. Comedones on the combined-SSA side were reduced more after six treatments, that the mean reduction rate of the combined-SSA side was 85.76%, and that of the CO2 laser-treated side was 62.32% (Pbetween < 0.001). Combining SSA also showed a better effect on reducing PI and TI than CO2 laser singly (Pbetween < 0.001). TEWL and HI between the two sides showed no significant differences after treatments. No permanent or severe side effects were observed on both side. CONCLUSIONS: The treatment combined CO2 laser with 30% SSA dealt with the efficacy-time conflict while significantly reducing comedones and improving skin texture in 12 weeks and no serious adverse reactions occurred. LIMITATIONS: It is a single-center study and the number of subjects was small.
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Droplet microfluidics is a rapidly advancing area of microfluidic technology, which offers numerous advantages for cell analysis, such as isolation and accumulation of signals, by confining cells within droplets. However, controlling cell numbers in droplets is challenging due to the uncertainty of random encapsulation which result in many empty droplets. Therefore, more precise control techniques are needed to achieve efficient encapsulation of cells within droplets. Here, an innovative microfluidic droplet manipulation platform had been developed, which employed positive pressure as a stable and controllable driving force for manipulating fluid within chips. The air cylinder, electro-pneumatics proportional valve, and the microfluidic chip were connected through a capillary, which enabled the formation of a fluid wall by creating a difference in hydrodynamic resistance between two fluid streams at the channel junction. Lowering the pressure of the driving oil phase eliminates hydrodynamic resistance and breaks the fluid wall. Regulating the duration of the fluid wall breakage controls the volume of the introduced fluid. Several important droplet microfluidic manipulations were demonstrated on this microfluidic platform, such as sorting of cells/droplets, sorting of droplets co-encapsulated cells and hydrogels, and active generation of droplets encapsulated with cells in a responsive manner. The simple, on-demand microfluidic platform was featured with high stability, good controllability, and compatibility with other droplet microfluidic technologies.
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Enteric pathogens are exposed to a dynamic polymicrobial environment in the gastrointestinal tract1. This microbial community has been shown to be important during infection, but there are few examples illustrating how microbial interactions can influence the virulence of invading pathogens2. Here we show that expansion of a group of antibiotic-resistant, opportunistic pathogens in the gut-the enterococci-enhances the fitness and pathogenesis of Clostridioides difficile. Through a parallel process of nutrient restriction and cross-feeding, enterococci shape the metabolic environment in the gut and reprogramme C. difficile metabolism. Enterococci provide fermentable amino acids, including leucine and ornithine, which increase C. difficile fitness in the antibiotic-perturbed gut. Parallel depletion of arginine by enterococci through arginine catabolism provides a metabolic cue for C. difficile that facilitates increased virulence. We find evidence of microbial interaction between these two pathogenic organisms in multiple mouse models of infection and patients infected with C. difficile. These findings provide mechanistic insights into the role of pathogenic microbiota in the susceptibility to and the severity of C. difficile infection.
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Clostridioides difficile , Enterococcus , Interacciones Microbianas , Animales , Humanos , Ratones , Antibacterianos/farmacología , Arginina/deficiencia , Arginina/metabolismo , Clostridioides difficile/metabolismo , Clostridioides difficile/patogenicidad , Clostridioides difficile/fisiología , Modelos Animales de Enfermedad , Farmacorresistencia Bacteriana , Enterococcus/efectos de los fármacos , Enterococcus/metabolismo , Enterococcus/patogenicidad , Enterococcus/fisiología , Microbioma Gastrointestinal/efectos de los fármacos , Intestinos/efectos de los fármacos , Intestinos/metabolismo , Intestinos/microbiología , Leucina/metabolismo , Ornitina/metabolismo , Virulencia , Susceptibilidad a EnfermedadesRESUMEN
Gastric carcinogenesis is mediated by complex interactions among Helicobacter pylori, host, and environmental factors. Here, we demonstrate that H. pylori augmented gastric injury in INS-GAS mice under iron-deficient conditions. Mechanistically, these phenotypes were not driven by alterations in the gastric microbiota; however, discovery-based and targeted metabolomics revealed that bile acids were significantly altered in H. pylori-infected mice with iron deficiency, with significant upregulation of deoxycholic acid (DCA), a carcinogenic bile acid. The severity of gastric injury was further augmented when H. pylori-infected mice were treated with DCA, and, in vitro, DCA increased translocation of the H. pylori oncoprotein CagA into host cells. Conversely, bile acid sequestration attenuated H. pylori-induced injury under conditions of iron deficiency. To translate these findings to human populations, we evaluated the association between bile acid sequestrant use and gastric cancer risk in a large human cohort. Among 416,885 individuals, a significant dose-dependent reduction in risk was associated with cumulative bile acid sequestrant use. Further, expression of the bile acid receptor transmembrane G protein-coupled bile acid receptor 5 (TGR5) paralleled the severity of carcinogenic lesions in humans. These data demonstrate that increased H. pylori-induced injury within the context of iron deficiency is tightly linked to altered bile acid metabolism, which may promote gastric carcinogenesis.
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Infecciones por Helicobacter , Helicobacter pylori , Deficiencias de Hierro , Neoplasias Gástricas , Animales , Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Ácidos y Sales Biliares/metabolismo , Carcinogénesis/metabolismo , Mucosa Gástrica/metabolismo , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/genética , Infecciones por Helicobacter/metabolismo , Helicobacter pylori/genética , Helicobacter pylori/metabolismo , Humanos , Inflamación/patología , Ratones , Neoplasias Gástricas/genéticaRESUMEN
Eukaryotic cells recognize intracellular pathogens through pattern recognition receptors, including sensors of aberrant nucleic acid structures. Sensors of double-stranded RNA (dsRNA) are known to detect replication intermediates of RNA viruses. It has long been suggested that annealing of mRNA from symmetrical transcription of both top and bottom strands of DNA virus genomes can produce dsRNA during infection. Supporting this hypothesis, nearly all DNA viruses encode inhibitors of dsRNA-recognition pathways. However, direct evidence that DNA viruses produce dsRNA is lacking. Contrary to dogma, we show that the nuclear-replicating DNA virus adenovirus (AdV) does not produce detectable levels of dsRNA during infection. In contrast, abundant dsRNA is detected within the nucleus of cells infected with AdV mutants defective for viral RNA processing. In the presence of nuclear dsRNA, the cytoplasmic dsRNA sensor PKR is relocalized and activated within the nucleus. Accumulation of viral dsRNA occurs in the late phase of infection, when unspliced viral transcripts form intron/exon base pairs between top and bottom strand transcripts. We propose that DNA viruses actively limit dsRNA formation by promoting efficient splicing and mRNA processing, thus avoiding detection and restriction by host innate immune sensors of pathogenic nucleic acids.
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Adenoviridae , Empalme del ARN , ARN Viral , Adenoviridae/genética , Adenoviridae/metabolismo , ARN Bicatenario/genética , ARN Bicatenario/metabolismo , ARN Mensajero/metabolismo , ARN Viral/genética , ARN Viral/metabolismoRESUMEN
The basic function of the blood-brain barrier (BBB) is to selectively regulate the infiltration of solutes from the circulating blood into the central nervous system (CNS). Impaired BBB activity is related to brain damage caused by stroke, traumatic injury, neurodegenerative diseases, etc. Comprised of a monolayer of endothelial cells, the integrity of the BBB is determined by the expression of tight junction proteins and the contractile activity of the perijunctional apical actomyosin ring. Irbesartan, an AT1R antagonist, has been widely used for the treatment of hypertension. However, the pharmacological function of Irbesartan in the balance of the BBB is still unknown. In the present study, we performed both in-vivo and in-vitro experiments using lipopolysaccharide (LPS) to explore the mechanism behind the protective effects of Irbesartan against the BBB impairment. The results of our mouse model study revealed that Irbesartan could reduce BBB permeability, restore the expression of Occludin, and suppress the expression of inflammatory mediators, including interleukin-6, monocyte chemoattractant protein-1, and intercellular adhesion molecule-1. Additionally, Irbesartan improved LPS-induced depressive-like behavior. In our in vitro experiments, human brain microvascular endothelial cells (HBMVECs) stimulated with LPS demonstrated decreased endothelial permeability and increased occludin expression in response to Irbesartan treatment. Importantly, we found that the protective effects of Irbesartan were mediated through the NF-κB/MLC/MLCK signaling pathway, as blockage of NF-κB abolished the effects of Irbesartan. Our findings provide a basis for further research into the neuroprotective mechanism of Irbesartan.
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Depresión/tratamiento farmacológico , Células Endoteliales/fisiología , Hipertensión/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Irbesartán/uso terapéutico , Microvasos/patología , Fármacos Neuroprotectores/uso terapéutico , Actomiosina/metabolismo , Animales , Conducta Animal/efectos de los fármacos , Barrera Hematoencefálica , Permeabilidad Capilar , Línea Celular , Células Endoteliales/efectos de los fármacos , Humanos , Interleucina-6/metabolismo , Irbesartán/farmacología , Lipopolisacáridos/inmunología , Ratones , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Fármacos Neuroprotectores/farmacología , Ocludina/metabolismo , Receptor de Angiotensina Tipo 1/metabolismo , Receptores CCR2/metabolismo , Transducción de Señal , Uniones Estrechas/metabolismoRESUMEN
Exosome and microRNAs (miRs) are implicated in ischemia/reperfusion (I/R) process. In this study, I/R mouse model was established, and exosomes derived from human umbilical cord mesenchymal stem cells (hUCMSCs) were isolated, identified, and injected to I/R mice to observe nerve injury and microglia M1 polarization. The differentially expressed genes in I/R microglia from databases were analyzed, and miRs differentially expressed in exosomes-treated microglia were analyzed by microarray. miR-26b-5p expression in hUCMSCs was intervened. Besides, microglia was extracted and co-cultured with SH-SY5Y or PC12 cells in oxygen-glucose deprivation/reperfusion (OGD/R) models to simulate I/R in vivo. Additionally, Toll-like receptor (TLR) activator GS-9620 was added to microglia. Exosomes alleviated nerve injury and inhibited M1 polarization in microglia. After I/R modeling, CH25H expression in microglia was upregulated but decreased after exosome treatment. miR-26b-5p was upregulated in microglia after exosome treatment and could target CH25H. Reduction in exosomal miR-26b-5p reversed the effects of hUCMSCs-exos on microglia. TLR pathway was activated in microglia after I/R but exosomes prevented its activation. Exosomal miR-26b-5p could repress M1 polarization of microglia by targeting CH25H to inactivate the TLR pathway, so as to relieve nerve injury after cerebral I/R. This investigation may offer new approaches for I/R treatment.
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Isquemia Encefálica/metabolismo , Polaridad Celular/genética , Exosomas/metabolismo , MicroARNs/metabolismo , Microglía/metabolismo , Daño por Reperfusión/metabolismo , Animales , Hipoxia de la Célula , Técnicas de Cocultivo , Modelos Animales de Enfermedad , Glucosa/deficiencia , Humanos , Células Madre Mesenquimatosas/metabolismo , Ratones , Ratones Endogámicos BALB C , MicroARNs/genética , Células PC12 , Ratas , Esteroide Hidroxilasas/metabolismo , TransfecciónRESUMEN
Designing a multifunctional theranostic nanoplatform with optional therapeutic strategies is highly desirable to select the most suitable therapeutic manners for the patient's cancer treatment. Among all shapes of silver materials, a silver nanoprism was reported to have great potential in photothermal therapy (PTT) owing to its strong surface plasmon resonance band in the near-infrared region. However, its instability in physicochemical environments and its severe toxicity confined its further application. To overcome this, herein, we demonstrated a silver prism-polydopamine (PDA) hybrid nanoplatform for tumor treatment with three therapeutic strategies. Specifically, the PDA coating endows the silver prism with excellent stability, high photothermal conversion, long-term in vivo biocompatibility, ease of decorating targeting ligands, and drug delivery. Upon near-infrared laser irradiation (808 nm, 1 W/cm2), tumors can be eradicated by the as-prepared nanoparticle through monomodal PTT. Besides, when combined with a chemical drug, this nanoparticle is able to inhibit tumor growth via combined photochemotherapy under a lower laser treatment (0.7 W/cm2). Furthermore, by supplementing with an immune checkpoint blockade, the realized synergistic photochemoimmunotherapy exhibits high efficacy to inhibit tumor relapse and metastasis. Moreover, owing to the high photothermal conversion efficiency and great X-ray attenuation ability of the silver nanoprism, our designed nanoplatform can be used in photoacoustic, computed tomography, and infrared thermal multimodal imaging. Our study provides a multifunctional nanoparticle for tumor theranostics, and this therapeutic strategy-optional nanoplatform shows promise in future biomedicine.
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Antineoplásicos , Nanopartículas del Metal/química , Imagen Multimodal/métodos , Fotoquimioterapia/métodos , Nanomedicina Teranóstica/métodos , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Apoptosis/efectos de la radiación , Línea Celular Tumoral , Sistemas de Liberación de Medicamentos , Femenino , Células Hep G2 , Humanos , Indoles/química , Rayos Infrarrojos , Ratones , Ratones Endogámicos BALB C , Polímeros/química , PlataRESUMEN
Stimulated cells and cancer cells have widespread shortening of mRNA 3'-untranslated regions (3'UTRs) and switches to shorter mRNA isoforms due to usage of more proximal polyadenylation signals (PASs) in introns and last exons. U1 snRNP (U1), vertebrates' most abundant non-coding (spliceosomal) small nuclear RNA, silences proximal PASs and its inhibition with antisense morpholino oligonucleotides (U1 AMO) triggers widespread premature transcription termination and mRNA shortening. Here we show that low U1 AMO doses increase cancer cells' migration and invasion in vitro by up to 500%, whereas U1 over-expression has the opposite effect. In addition to 3'UTR length, numerous transcriptome changes that could contribute to this phenotype are observed, including alternative splicing, and mRNA expression levels of proto-oncogenes and tumor suppressors. These findings reveal an unexpected role for U1 homeostasis (available U1 relative to transcription) in oncogenic and activated cell states, and suggest U1 as a potential target for their modulation.
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Movimiento Celular , Neoplasias/metabolismo , Ribonucleoproteína Nuclear Pequeña U1/metabolismo , Línea Celular Tumoral , Humanos , Invasividad Neoplásica , Neoplasias/genética , Neoplasias/patología , Neoplasias/fisiopatología , Oligonucleótidos Antisentido/genética , Oligonucleótidos Antisentido/metabolismo , Empalme del ARN , ARN Mensajero/metabolismo , Ribonucleoproteína Nuclear Pequeña U1/genéticaRESUMEN
Ankle joint kinematics is mainly stabilized by the morphology of the talar dome and the articular surface of tibiofibular mortise as well as the medial and lateral ligament complexes. Because of this the bicondylar geometry of talus dome is believed to be crucial for ankle implant design. However, little data exist describing the precise anatomy of the talar dome and the talocrural joint axis. The aim of this study is to document the anatomy of the talar dome and the axis of the talocrural joint using three-dimensional (3D) computed tomographic (CT) modeling. Seventy-one participants enrolled for CT scanning and 3D talar model reconstruction. All the ankles were held in a neutral position during the CT scanning. Six points on the lateral and medial crest of the talar dome were defined. The coordinate of the six points; radii of lateral-anterior (R-LA), lateral-posterior (R-LP), medial-anterior (R-MA), and medial-posterior (R-MP) sections; and inclination angle of the talar dome were measured, and the inclination and deviation angles of the talocrural joint axis were determined. The mean values of R-LA, R-LP, R-MA, and R-MP were 19.23 ± 2.47 mm, 18.76 ± 2.90 mm, 17.02 ± 3.49 mm, and 22.75 ± 3.04 mm. The mean inclination angle of the talar dome was 9.86 ± 3.30 degrees. Gender variation was found in this parameter. The mean inclination and deviation angles were 8.60 ± 0.07 and 0.76 ± 0.69 degrees for the dorsiflexion axis and -7.34 ± 0.07 and 0.09 ± 0.18 degrees for the plantarflexion axis. Bilateral asymmetries between the medial and lateral crest of the talar dome were found, which resulted in different dorsiflexion and plantarflexion axes of the talocrural joint. Currently, no ankle implants replicate this talar anatomy, and these findings should be considered in future implant designs.
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Articulación del Tobillo , Imagenología Tridimensional , Prótesis Articulares , Diseño de Prótesis , Astrágalo , Tomografía Computarizada por Rayos X , Articulación del Tobillo/diagnóstico por imagen , Articulación del Tobillo/fisiopatología , Fenómenos Biomecánicos , Femenino , Humanos , Masculino , Astrágalo/diagnóstico por imagen , Astrágalo/fisiopatologíaRESUMEN
Full-length transcription in the majority of human genes depends on U1 snRNP (U1) to co-transcriptionally suppress transcription-terminating premature 3' end cleavage and polyadenylation (PCPA) from cryptic polyadenylation signals (PASs) in introns. However, the mechanism of this U1 activity, termed telescripting, is unknown. Here, we captured a complex, comprising U1 and CPA factors (U1-CPAFs), that binds intronic PASs and suppresses PCPA. U1-CPAFs are distinct from U1-spliceosomal complexes; they include CPA's three main subunits, CFIm, CPSF, and CstF; lack essential splicing factors; and associate with transcription elongation and mRNA export complexes. Telescripting requires U1:pre-mRNA base-pairing, which can be disrupted by U1 antisense oligonucleotide (U1 AMO), triggering PCPA. U1 AMO remodels U1-CPAFs, revealing changes, including recruitment of CPA-stimulating factors, that explain U1-CPAFs' switch from repressive to activated states. Our findings outline this U1 telescripting mechanism and demonstrate U1's unique role as central regulator of pre-mRNA processing and transcription.
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Núcleo Celular/metabolismo , Factor de Especificidad de Desdoblamiento y Poliadenilación/metabolismo , División del ARN , Precursores del ARN/biosíntesis , ARN Mensajero/biosíntesis , Ribonucleoproteína Nuclear Pequeña U1/metabolismo , Transcripción Genética , Regiones no Traducidas 3' , Transporte Activo de Núcleo Celular , Sitios de Unión , Núcleo Celular/genética , Factor de Especificidad de Desdoblamiento y Poliadenilación/genética , Factor de Estimulación del Desdoblamiento/genética , Factor de Estimulación del Desdoblamiento/metabolismo , Células HeLa , Humanos , Complejos Multiproteicos , Poli A/metabolismo , Unión Proteica , Precursores del ARN/genética , ARN Mensajero/genética , Ribonucleoproteína Nuclear Pequeña U1/genéticaRESUMEN
Recent observations showed that nascent RNA polymerase II transcripts, pre-mRNAs, and noncoding RNAs are highly susceptible to premature 3'-end cleavage and polyadenylation (PCPA) from numerous intronic cryptic polyadenylation signals (PASs). The importance of this in gene regulation was not previously appreciated as PASs, despite their prevalence, were thought to be active in terminal exons at gene ends. Unexpectedly, antisense oligonucleotide interference with U1 snRNA base-pairing to 5' splice sites, which is necessary for U1 snRNP's (U1) function in splicing, caused widespread PCPA in metazoans. This uncovered U1's PCPA suppression activity, termed telescripting, as crucial for full-length transcription in thousands of vertebrate genes, providing a general role in transcription elongation control. Progressive intron-size expansion in metazoan evolution greatly increased PCPA vulnerability and dependence on U1 telescripting. We describe how these observations unfolded and discuss U1 telescripting's role in shaping the transcriptome.
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Regulación de la Expresión Génica/fisiología , ARN Nuclear Pequeño/metabolismo , Ribonucleoproteínas Nucleares Pequeñas/metabolismo , Terminación de la Transcripción Genética , Animales , Humanos , Intrones , ARN Nuclear Pequeño/genéticaRESUMEN
Telescripting is a fundamental cotranscriptional gene regulation process that relies on U1 snRNP (U1) to suppress premature 3'-end cleavage and polyadenylation (PCPA) in RNA polymerase II (Pol II) transcripts, which is necessary for full-length transcription of thousands of protein-coding (pre-mRNAs) and long noncoding (lncRNA) genes. Like U1 role in splicing, telescripting requires U1 snRNA base-pairing with nascent transcripts. Inhibition of U1 base-pairing with U1 snRNA antisense morpholino oligonucleotide (U1 AMO) mimics widespread PCPA from cryptic polyadenylation signals (PASs) in human tissues, including PCPA in introns and last exons' 3'-untranslated regions (3' UTRs). U1 telescripting-PCPA balance changes generate diverse RNAs depending on where in a gene it occurs. Long genes are highly U1-telescripting-dependent because of PASs in introns compared to short genes. Enrichment of cell cycle control, differentiation, and developmental functions in long genes, compared to housekeeping and acute cell stress response genes in short genes, reveals a gene size-function relationship in mammalian genomes. This polarization increased in metazoan evolution by previously unexplained intron expansion, suggesting that U1 telescripting could shift global gene expression priorities. We show that that modulating U1 availability can profoundly alter cell phenotype, such as cancer cell migration and invasion, underscoring the critical role of U1 homeostasis and suggesting it as a potential target for therapies. We describe a complex of U1 with cleavage and polyadenylation factors that silences PASs in introns and 3' UTR, which gives insights into U1 telescripting mechanism and transcription elongation regulation.
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Acoustic bottle-beams have been realized by acoustic metasurfaces (AMs) composed of space-coiling subunits. By manipulating the transmitted acoustical phase, the special AM can generate two intersecting accelerating beams along the designed convex trajectories, forming the acoustic bottle-beam. The transmitted acoustic bottle-beams are investigated theoretically and demonstrated numerically. We find that the shape and area of the acoustic bottle-beam could be statically controlled by designing the AM as well as dynamically modulated by the incident angles. In addition, the highly efficient acoustic focusing could be obtained at the convergence point of the bottle-beams, which also could be adjusted dynamically by the incident angles. It is further found that this focusing is robust against the obstacle scattering. The realization and manipulation of acoustic bottle-beams may have potential applications in biomedical imaging/therapy and non-destructive evaluation.
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Understanding the shape of superior talar dome is essential for a better size compatibility between talar component of ankle implant and bone. The purpose of this study was to determine whether there were gender variations in (1) width (TW) and length (TL) of talus, as well as anterior width (DAW), middle width (DMW), posterior width (DPW), and length (DL) of superior talar dome; (2) differences between the DAW, DMW, and DPW; (3) the ratios between these parameters. Fifty-one cadaveric ankle specimens were included. Two observers measured all the specimens using vernier caliper. Intraclass correlation coefficients (ICCs) were used for intraobserver and interobserver reliability analysis and the reliability was thought to be good if the ICC>0.75. A two-tailed unpaired t-test or the rank-sum test was used to investigate gender variations. A single-factor ANOVA was utilized to identify the differences between the width of the superior talar dome surface and p value of <0.05 was considered significant. Intraobserver and interobserver reliability were good. Significant gender variations were found, in which TW, TL, DAW, DMW, DPW, and DL of female specimens were much smaller than those of male. The width of talar dome linearly decreased from DAW to DPW; however, the linearly decreased rate from anterior to posterior width was bigger in female. Moreover, significant differences were found in DAW/DPW, DMW/DPW, DL/DAW, DL/DMW, and DL/DPW between male and female. Based on our result, there was no difference in the 2D shape of the whole talus instead gender variation existed in the 2D shape of superior talar dome between male and female. The current 2D data could contribute to figure out more suitable size of talar component for Chinese population and might indicate a gender-specific shape of bone-implant interface, which could reduce the potential bone-component incompatibility when performing ankle replacement using standard component.
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Astrágalo/anatomía & histología , Articulación del Tobillo , Cadáver , China , Femenino , Humanos , Masculino , Reproducibilidad de los Resultados , Caracteres SexualesRESUMEN
U1 snRNP (U1) functions in splicing introns and telescripting, which suppresses premature cleavage and polyadenylation (PCPA). Using U1 inhibition in human cells, we show that U1 telescripting is selectively required for sustaining long-distance transcription elongation in introns of large genes (median 39 kb). Evidence of widespread PCPA in the same locations in normal tissues reveals that large genes incur natural transcription attrition. Underscoring the importance of U1 telescripting as a gene-size-based mRNA-regulation mechanism, small genes were not sensitive to PCPA, and the spliced-mRNA productivity of â¼1,000 small genes (median 6.8 kb) increased upon U1 inhibition. Notably, these small, upregulated genes were enriched in functions related to acute stimuli and cell-survival response, whereas genes subject to PCPA were enriched in cell-cycle progression and developmental functions. This gene size-function polarization increased in metazoan evolution by enormous intron expansion. We propose that telescripting adds an overarching layer of regulation to size-function-stratified genomes, leveraged by selective intron expansion to rapidly shift gene expression priorities.
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Regulación de la Expresión Génica , Genoma Humano , Poliadenilación , Ribonucleoproteína Nuclear Pequeña U1/metabolismo , Transcripción Genética , HumanosRESUMEN
Long noncoding RNAs (lncRNAs), a recently discovered class of cellular RNAs, play important roles in the regulation of many cellular developmental processes. Although lncRNAs have been systematically identified in various systems, most of them have not been functionally characterized in vivo in animal models. In this study, we identified 128 testis-specific Drosophila lncRNAs and knocked out 105 of them using an optimized three-component CRISPR/Cas9 system. Among the lncRNA knockouts, 33 (31%) exhibited a partial or complete loss of male fertility, accompanied by visual developmental defects in late spermatogenesis. In addition, six knockouts were fully or partially rescued by transgenes in a trans configuration, indicating that those lncRNAs primarily work in trans Furthermore, gene expression profiles for five lncRNA mutants revealed that testis-specific lncRNAs regulate global gene expression, orchestrating late male germ cell differentiation. Compared with coding genes, the testis-specific lncRNAs evolved much faster. Moreover, lncRNAs of greater functional importance exhibited higher sequence conservation, suggesting that they are under constant evolutionary selection. Collectively, our results reveal critical functions of rapidly evolving testis-specific lncRNAs in late Drosophila spermatogenesis.
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Secuencia Conservada/genética , ARN Largo no Codificante/genética , Espermatogénesis/genética , Testículo/crecimiento & desarrollo , Animales , Sistemas CRISPR-Cas , Drosophila/genética , Drosophila/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Células Germinativas/crecimiento & desarrollo , Infertilidad Masculina/genética , Infertilidad Masculina/patología , MasculinoRESUMEN
BACKGROUND: Recently, many lncRNAs were found to be deregulated in various human cancers and play important roles in carcinogenesis. MATERIAL AND METHODS: To investigate the association of lncRNAs to gastrointestinal cancers, 12 cases of esophageal cancer and hepatic cancer, 16 cases of gastric cancer and colorectal cancer and their matched adjacent tissue samples, 12 esophageal cancer cell lines, 7 gastric cancer cell lines, 10 colorectal cancer cell lines, and 11 hepatic cancer cell lines were examined. RNA-seq, bioinformatics pipeline, co-expression network, and gene ontology enrichment analysis were performed. RESULTS: We have identified 23,169 candidate novel lncRNA transcripts. Comparing with protein coding genes the lncRNAs tend to be shorter and have less exons. Remarkably, we found 15 lncRNAs that were down-regulated in all cancer cell lines among all four types of cancers. In addition, we analyzed the differentially expressed lncRNAs in gastrointestinal cancer cells before and after treatment with 5-Aza. Many lncRNAs were up-regulated after 5-Aza treatment, which suggested that the expression of these lncRNAs may be regulated by DNA methylation. Finally, based on the co-expression network and GO enrichment analysis, we predicted that many novel lncRNAs were involved in pathways like apoptosis, cell cycle, cell adhesion, EMT, epigenetic regulation, DNA damage response signaling, and immune response. CONCLUSION: Based on RNA-Seq and bioinformatics analysis, we have identified a significant number of novel lncRNAs, which could be involved in important pathways related to gastrointestinal cancer development. Overall, we provide a rich resource for identifying new biomarkers and studying novel lncRNA regulation pathways in gastrointestinal cancer.
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Carcinogénesis/genética , Neoplasias Gastrointestinales/genética , Regulación Neoplásica de la Expresión Génica , ARN Largo no Codificante/genética , Transducción de Señal/genética , Apoptosis/genética , Azacitidina/farmacología , Biomarcadores de Tumor/genética , Ciclo Celular/genética , Línea Celular Tumoral , Metilación de ADN , Regulación hacia Abajo , Epigénesis Genética , Transición Epitelial-Mesenquimal/genética , Perfilación de la Expresión Génica , Humanos , ARN Largo no Codificante/efectos de los fármacos , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de Secuencia de ARN , Regulación hacia ArribaRESUMEN
JASMONATE ZIM-domain (JAZ) proteins play important roles in plant defence and growth by regulating jasmonate signalling. Through data mining, we discovered that the JAZ7 gene was up-regulated in darkness. In the dark, the jaz7 mutant displayed more severe leaf yellowing, quicker chlorophyll degradation, and higher hydrogen peroxide accumulation compared with wild-type (WT) plants. The mutant phenotype of dark-induced leaf senescence could be rescued in the JAZ7-complemented and -overexpression lines. Moreover, the double mutants of jaz7 myc2 and jaz7 coi1 exhibited delayed leaf senescence. We further employed GeneChip analysis to study the molecular mechanism. Some key genes down-regulated in the triple mutant myc2 myc3 myc4 were up-regulated in the jaz7 mutant under darkness. The Gene Ontology terms 'leaf senescence' and 'cell death' were significantly enriched in the differentially expressed genes. Combining the genetic and transcriptomic analyses together, we proposed a model whereby darkness can induce JAZ7, which might further block MYC2 to suppress dark-induced leaf senescence. In darkness, the mutation of JAZ7 might partially liberate MYC2/MYC3/MYC4 from suppression, leading the MYC proteins to bind to the G-box/G-box-like motifs in the promoters, resulting in the up-regulation of the downstream genes related to indole-glucosinolate biosynthesis, sulphate metabolism, callose deposition, and JA-mediated signalling pathways. In summary, our genetic and transcriptomic studies established the JAZ7 protein as an important regulator in dark-induced leaf senescence.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Oscuridad , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/metabolismo , Proteínas Represoras/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes de Plantas , Peróxido de Hidrógeno/farmacología , Modelos Biológicos , Mutación/genética , Fenotipo , Hojas de la Planta/efectos de los fármacos , Plantas Modificadas Genéticamente , Unión Proteica/efectos de los fármacos , Proteínas Represoras/genética , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transcriptoma/genéticaRESUMEN
OBJECTIVE: To investigate the effect of ligation level of inferior mesenteric artery (IMA) on postoperative defecation function in patients with rectal cancer. METHODS: A total of 128 rectal cancer patients who were planned to undergo low anterior resection in the First Hospital of Zibo City between January 1, 2012 and December 31, 2013 were prospectively enrolled and randomly divided into IMA high ligation group(63 cases, cutting distance of 1.0 to 1.5 cm to the root of IMA) and low ligation group(65 cases, cutting distance of 0.5 to 1.0 cm to the root of left colic artery originated from IMA). The efficacy, especially the defecation function, was observed and compared 3 months and 1 year after surgery between the two groups. RESULTS: No significant difference was found in the number of harvested lymph nodes between two groups[8(1-30) vs. 7(2-28), P=0.125], but high ligation group had greater number of metastatic lymph nodes[1(0-9) vs. 0(0-8), P=0.041]. Frequency of defecation in high ligation group was significantly higher than that in low ligation group during postoperative 3-month follow-up[5(2-10)/d vs. 3(1-8)/d, P=0.035], whereas other indexes of defecation function were not significantly different(all P>0.05). The proportion of patients needing laxatives in high ligation group was higher than that in low ligation group during postoperative 1-year follow-up [11.3%(6/53) vs. 1.7%(1/58), P=0.038], whereas other indexes of defecation function were not significantly different as well (all P>0.05). Three cases and 2 cases showed recurrence in high ligation group and low ligation group respectively during postoperative 1-year follow-up without significant difference(P=0.623). CONCLUSION: Low ligation of IMA in low anterior resection for rectal cancer is beneficial to the protection against defecation function.