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The construction of efficient and low toxic non-viral gene delivery vectors is of great significance for gene therapy. Herein, two novel polycations were constructed via Michael addition from low molecular weight polyethylenimine (PEI) 600 Da and amino acid-containing linkages. Lysine and histidine were introduced for the purpose of improved DNA binding and pH buffering capacity, respectively. The ester bonds afforded the polymer biodegradability, which was confirmed by the gel permeation chromatography (GPC) measurement. The polymers could well condense DNA into nanoparticles and protect DNA from degradation by nuclease. Compared with PEI 25 kDa, these polymers showed higher transfection efficiency, lower toxicity, and better serum tolerance. Study of this mechanism revealed that the polyplexes enter the cells mainly through caveolae-mediated endocytosis pathway; this, together with their biodegradability, facilitates the internalization of polyplexes and the release of DNA. The results reveal that the amino acid-linked low molecular weight PEI polymers could serve as promising candidates for non-viral gene delivery.
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Aminoácidos/química , ADN/química , Nanopartículas/química , Polietileneimina/química , Aminoácidos/genética , Aminoácidos/uso terapéutico , Materiales Biocompatibles/química , Materiales Biocompatibles/uso terapéutico , ADN/genética , ADN/uso terapéutico , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/genética , Endocitosis/efectos de los fármacos , Técnicas de Transferencia de Gen/tendencias , Terapia Genética/métodos , Humanos , Peso Molecular , Nanopartículas/uso terapéutico , Plásmidos/genética , Polietileneimina/uso terapéutico , Polímeros/químicaRESUMEN
Polyvinyl alcohol (PVA) is used widely in industry, and associated environmental pollution is a serious problem. Herein, we report a novel, efficient PVA degrader, Stenotrophomonas rhizophila QL-P4, isolated from fallen leaves from a virgin forest in the Qinling Mountains. The complete genome was obtained using single-molecule real-time (SMRT) technology and corrected using Illumina sequencing. Bioinformatics analysis revealed eight PVA/vinyl alcohol oligomer (OVA)-degrading genes. Of these, seven genes were predicted to be involved in the classic intracellular PVA/OVA degradation pathway, and one (BAY15_3292) was identified as a novel PVA oxidase. Five PVA/OVA-degrading enzymes were purified and characterized. One of these, BAY15_1712, a PVA dehydrogenase (PVADH), displayed high catalytic efficiency toward PVA and OVA substrate. All reported PVADHs only have PVA-degrading ability. Most importantly, we discovered a novel PVA oxidase (BAY15_3292) that exhibited higher PVA-degrading efficiency than the reported PVADHs. Further investigation indicated that BAY15_3292 plays a crucial role in PVA degradation in S. rhizophila QL-P4. Knocking out BAY15_3292 resulted in a significant decline in PVA-degrading activity in S. rhizophila QL-P4. Interestingly, we found that BAY15_3292 possesses exocrine activity, which distinguishes it from classic PVADHs. Transparent circle experiments further proved that BAY15_3292 greatly affects extracellular PVA degradation in S. rhizophila QL-P4. The exocrine characteristics of BAY15_3292 facilitate its potential application to PVA bioremediation. In addition, we report three new efficient secondary alcohol dehydrogenases (SADHs) with OVA-degrading ability in S. rhizophila QL-P4; in contrast, only one OVA-degrading SADH was reported previously.IMPORTANCE With the widespread application of PVA in industry, PVA-related environmental pollution is an increasingly serious issue. Because PVA is difficult to degrade, it accumulates in aquatic environments and causes chronic toxicity to aquatic organisms. Biodegradation of PVA, as an economical and environment-friendly method, has attracted much interest. To date, effective and applicable PVA-degrading bacteria/enzymes have not been reported. Herein, we report a new efficient PVA degrader (S. rhizophila QL-P4) that has five PVA/OVA-degrading enzymes with high catalytic efficiency, among which BAY15_1712 is the only reported PVADH with both PVA- and OVA-degrading abilities. Importantly, we discovered a novel PVA oxidase (BAY15_3292) that is not only more efficient than other reported PVA-degrading PVADHs but also has exocrine activity. Overall, our findings provide new insight into PVA-degrading pathways in microorganisms and suggest S. rhizophila QL-P4 and its enzymes have the potential for application to PVA bioremediation to reduce or eliminate PVA-related environmental pollution.
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Proteínas Bacterianas/genética , Genoma Bacteriano , Alcohol Polivinílico/metabolismo , Stenotrophomonas/genética , Stenotrophomonas/metabolismo , Proteínas Bacterianas/metabolismo , Biología Computacional , Alineación de Secuencia , Análisis de Secuencia de ADN , Stenotrophomonas/enzimologíaRESUMEN
In this study, the photochemical internalization (PCI) technique was adopted in a nanoparticle-based antigen delivery system to enhance antigen-specific CD8+ T cell immune response for cancer immunotherapy. Pheophorbide A, a hydrophobic photosensitizer, grafted with polyethylenimine (PheoA-PEI) with endosome escape activity and near-infrared imaging capability was prepared. A model antigen ovalbumin (OVA) was then complexed with PheoA-PEI to form PheoA-PEI/OVA nanoparticles (PheoA-PEI/OVA NPs) that are responsive to light. Flow cytometry analysis revealed increased endocytosis in a murine dendritic cell line (DC2.4) that was treated with PheoA-PEI/OVA NPs compared to free OVA. Generation of reactive oxygen species (ROS) in DC2.4 cells was also confirmed quantitatively and qualitatively using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). Confocal laser scanning microscopy (CLSM) further demonstrated that the PheoA-PEI/OVA NPs enhanced cytosolic antigen release after light stimulation. Moreover, PheoA-PEI/OVA NP treated DC2.4 cells exhibited enhanced cross-presentation to B3Z T cell hybridoma in vitro after light irradiation, substantially increased compared to those treated with free OVA. Consistently, in vivo results revealed upregulation of CD3+CD8+T lymphocytes in tumors of mice treated with dendritic cells plus PheoA-PEI/OVA NPs and light irradiation. The activated T cell response is partly responsible for the inhibitory effect on E.G7 tumor growth in mice immunized with dendritic cells plus PheoA-PEI/OVA NPs and light irradiation. Our results demonstrate the feasibility to enhance antigen-specific CD8+ T cell immune response by light-responsive nanoparticle-based vaccine delivery for cancer immunotherapy.
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Clorofila/análogos & derivados , Células Dendríticas/metabolismo , Inmunoterapia/métodos , Nanopartículas/química , Polietileneimina/química , Animales , Linfocitos T CD8-positivos/metabolismo , Línea Celular , Línea Celular Tumoral , Clorofila/química , Femenino , Citometría de Flujo , Ratones , Ratones Endogámicos C57BL , Microscopía Confocal , Especies Reactivas de Oxígeno/metabolismo , Oxígeno Singlete/metabolismoRESUMEN
OBJECTIVES: To study the microbial community structure on the root surface of patients with periodontitis. METHODS: Bacterial plaque and tissues from the root neck (RN group),root middle (RM group) and root tine (RT group) of six teeth with mobility 3 in one patient with periodontitis were sampled.The V3V4 region of 16S rRNA was sequenced on the Illumina MiSeq platform.The microbial community structure was analyzed by Mothur,Qiime and SPSS software. RESULTS: The principal component analysis (PCoA) results indicated that the RM samples had a similar microbial community structure as that of the RT samples,which was significant different from that of the RN samples.Thirteen phyla were detected in the three groups of samples,which included 7 dominant phyla.29 dominant genera were detected in 184 genera.The abundance of Bacteroidetes_[G-6] and Peptostre ptococcaceae_[XI][G-4] had a positive correlation with the depth of the collection site of samples (P<0.05),while the abundance of Prevotella,Selenomonas,Corynebacterium and Olsenella had a negative correlation with the depth of the collection site of samples (P<0.05). CONCLUSIONS: There is region-specificity of microbial community structure on the root surface of patients with periodontitis.
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Bacterias/clasificación , Placa Dental/microbiología , Periodontitis/microbiología , Raíz del Diente/microbiología , Humanos , ARN Ribosómico 16S/genéticaRESUMEN
Nanoscale drug delivery platforms have been developed over the past four decades that have shown promising clinical results in several types of cancer and inflammatory disorders. These nanocarriers carrying therapeutic payloads are maximizing the therapeutic outcomes while minimizing adverse effects. Yet one of the major challenges facing drug developers is the dilemma of premature versus on-demand drug release, which influences the therapeutic regiment, efficacy and potential toxicity. Herein, we report on redox-sensitive polymer-drug conjugate micelles for on-demand intracellular delivery of a model active agent, curcumin. Biodegradable methoxy poly(ethylene glycol)-poly(lactic acid) copolymer (mPEG-PLA) was conjugated with curcumin via a disulfide bond or ester bond (control), respectively. The self-assembled redox-sensitive micelles exhibited a hydrodynamic size of 115.6 ± 5.9 (nm) with a zeta potential of -10.6 ± 0.7 (mV). The critical micelle concentration was determined at 6.7 ± 0.4 (µg mL(-1)). Under sink conditions with a mimicked redox environment (10 mM dithiothreitol), the extent of curcumin release at 48 h from disulfide bond-linked micelles was nearly three times higher compared to the control micelles. Such rapid release led to a lower half maximal inhibitory concentration (IC50) in HeLa cells at 18.5 ± 1.4 (µg mL(-1)), whereas the IC50 of control micelles was 41.0 ± 2.4 (µg mL(-1)). The cellular uptake study also revealed higher fluorescence intensity for redox-sensitive micelles. In conclusion, the redox-sensitive polymeric conjugate micelles could enhance curcumin delivery while avoiding premature release, and achieving on-demand release under the high glutathione concentration in the cell cytoplasm. This strategy opens new avenues for on-demand drug release of nanoscale intracellular delivery platforms that ultimately might be translated into pre-clinical and future clinical practice.
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Portadores de Fármacos , Micelas , Nanotecnología/métodos , Materiales Biocompatibles/química , Dominio Catalítico , Supervivencia Celular , Curcumina/química , Citoplasma/metabolismo , Disulfuros/química , Glutatión/química , Células HeLa , Humanos , Hidrodinámica , Concentración de Iones de Hidrógeno , Inflamación , Concentración 50 Inhibidora , Lactatos/química , Espectroscopía de Resonancia Magnética , Microscopía Electrónica de Transmisión , Peso Molecular , Nanomedicina/métodos , Nanopartículas/química , Oxidación-Reducción , Tamaño de la Partícula , Polietilenglicoles/química , Polímeros/química , TemperaturaRESUMEN
Poor aqueous solubility, potential degradation, rapid metabolism and elimination lead to low bioavailability of pleiotropic impotent curcumin. Herein, we report two types of acid-responsive polymeric micelles where curcumin was encapsulated via both covalent and non-covalent modes for enhanced loading capacity and on-demand release. Biodegradable methoxy poly(ethylene glycol)-poly(lactic acid) copolymer (mPEG-PLA) was conjugated with curcumin via a hydrazone linker, generating two conjugates differing in architecture (single-tail versus double-tail) and free curcumin was encapsulated therein. The two micelles exhibited similar hydrodynamic size at 95 ± 3 nm (single-tail) and 96 ± 3 nm (double-tail), but their loading capacities differed significantly at 15.0 ± 0.5% (w/w) (single-tail) and 4.8 ± 0.5% (w/w) (double-tail). Under acidic sink conditions (pH 5.0 and 6.0), curcumin displayed a faster release from the single-tail nanocarrier, which was correlated to a low IC50 of 14.7 ± 1.6 (µg mL(-1)) compared to the value of double-tail micelle (24.9 ± 1.3 µg mL(-1)) in HeLa cells. The confocal imaging and flow cytometry analysis demonstrated a superior capability of single-tail micelle for intracellular curcumin delivery, which was a consequence of the higher loading capacity and lower degree of mPEG surface coverage. In conclusion, the dual loading mode is an effective means to increase the drug content in the micellar nanocarriers whose delivery efficiency is highly dependent on its polymer-drug conjugate architecture. This strategy offers an alternative nanoplatform for intracellularly delivering impotent hydrophobic agents (i.e. curcumin) in an efficient stimuli-triggered way, which is valuable for the enhancement of curcumin's efficacy in managing a diverse range of disorders.
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Supervivencia Celular/efectos de los fármacos , Curcumina/química , Portadores de Fármacos/química , Lactatos/química , Micelas , Nanopartículas/química , Polietilenglicoles/química , Curcumina/farmacología , Células HeLa , HumanosRESUMEN
OBJECTIVE: To study microbial diversity of peri-implantitis subgingival with high-throughput sequencing, and investigate microbiological etiology of peri-implantitis. METHODS: Subgingival plaques were sampled from the patients with peri-implantitis (D group) and non-peri-implantitis subjects (N group). The microbiological diversity of the subgingival plaques was detected by sequencing V4 region of 16S rRNA with Illumina Miseq platform. The diversity of the community structure was analyzed using Mothur software. RESULTS: A total of 156 507 gene sequences were detected in nine samples and 4 402 operational taxonomic units (OTUs) were found. Selenomonas, Pseudomonas, and Fusobacterium were dominant bacteria in D group, while Fusobacterium, Veillonella and Streptococcus were dominant bacteria in N group. Differences between peri-implantitis and non-peri-implantitis bacterial communities were observed at all phylogenetic levels by LEfSe, which was also found in PcoA test. CONCLUSION: The occurrence of peri-implantitis is not only related to periodontitis pathogenic microbe, but also related with the changes of oral microbial community structure. Treponema, Herbaspirillum, Butyricimonas and Phaeobacte may be closely related to the occurrence and development of peri-implantitis.
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Bacterias/clasificación , Placa Dental/microbiología , Periimplantitis/microbiología , ADN Bacteriano/genética , Fusobacterium , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Periodontitis , Filogenia , Pseudomonas , ARN Ribosómico 16S/genética , Selenomonas , Análisis de Secuencia de ADN , Streptococcus , TreponemaRESUMEN
Periodontitis is a common oral disease accompanied by inflammatory bone loss. The pathological characteristics of periodontitis usually accompany an imbalance in the periodontal immune microenvironment, leading to difficulty in bone regeneration. Therefore, effective treatment strategies are needed to modulate the immune environment in order to treat periodontitis. Here, highly-oriented periodic lamellae poly(ε-caprolactone) electrospun nanofibers (PLN) are developed by surface-directed epitaxial crystallization. The in vitro result shows that the PLN can precisely modulate macrophage polarization toward the M2 phenotype. Macrophages polarized by PLN significantly enhance the migration and osteogenic differentiation of Bone marrow stromal cells. Notably, results suggest that the topographical cues presented by PLN can modulate macrophage polarization by activating YAP, which reciprocally inhibits the NF-κB signaling pathway. The in vivo results indicate that PLN can inhibit inflammatory bone loss and facilitate bone regeneration in periodontitis. The authors' findings suggest that topographical nanofibers with periodic lamellae is a promising strategy for modulating immune environment to treat inflammatory bone loss in periodontitis.
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Nanofibras , Osteogénesis , Periodontitis , Poliésteres , Nanofibras/química , Periodontitis/terapia , Periodontitis/patología , Periodontitis/inmunología , Periodontitis/tratamiento farmacológico , Animales , Ratones , Poliésteres/química , Osteogénesis/efectos de los fármacos , Células RAW 264.7 , Macrófagos/inmunología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Regeneración Ósea/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , FN-kappa B/metabolismo , Células Madre Mesenquimatosas/inmunología , Inmunomodulación/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas Adaptadoras Transductoras de Señales/inmunología , Ratones Endogámicos C57BL , Masculino , Inflamación/patología , Proteínas Señalizadoras YAPRESUMEN
Herein, a series of biocompatible, robust, pH/sugar-sensitive, core-cross-linked, polyion complex (PIC) micelles based on phenylboronic acid-catechol interaction were developed for protein intracellular delivery. The rationally designed poly(ethylene glycol)-b-poly(glutamic acid-co-glutamicamidophenylboronic acid) (PEG-b-P(Glu-co-GluPBA)) and poly(ethylene glycol)-b-poly(l-lysine-co-ε-3,4-dihydroxyphenylcarboxyl-L-lysine) (PEG-b-P(Lys-co-LysCA)) copolymers were successfully synthesized and self-assembled under neutral aqueous condition to form uniform micelles. These micelles possessed a distinct core-cross-linked core-shell structure comprised of the PEG outer shell and the PGlu/PLys polyion complex core bearing boronate ester cross-linking bonds. The cross-linked micelles displayed superior physiological stabilities compared with their non-cross-linked counterparts while swelling and disassembling in the presence of excess fructose or at endosomal pH. Notably, either negatively or positively charged proteins can be encapsulated into the micelles efficiently under mild conditions. The in vitro release studies showed that the release of protein cargoes under physiological conditions was minimized, while a burst release occurred in response to excess fructose or endosomal pH. The cytotoxicity of micelles was determined by cck-8 assay in HepG2 cells. The cytochrome C loaded micelles could efficiently delivery proteins into HepG2 cells and exhibited enhanced apoptosis ability. Hence, this type of core-cross-linked PIC micelles has opened a new avenue to intracellular protein delivery.
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Antineoplásicos/farmacología , Reactivos de Enlaces Cruzados/química , Citocromos c/metabolismo , Fructosa/farmacología , Micelas , Polímeros/química , Antineoplásicos/síntesis química , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Ácidos Borónicos/química , Ácidos Borónicos/metabolismo , Catecoles/química , Catecoles/metabolismo , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Células Hep G2 , Humanos , Concentración de Iones de Hidrógeno , Iones/química , Iones/farmacología , Estructura Molecular , Relación Estructura-ActividadRESUMEN
Objective: The development of ankylosing spondylitis (AS) is closely related to autoimmune system dysfunction. Type 1 diabetes mellitus (T1DM) is an autoimmune disease that is a risk factor for many diseases. This study aimed to investigate the causal relationship between T1DM mellitus and AS genetically. Methods: A genome-wide association study (GWAS) of causal relationships between exposure (T1DM) and outcome (AS) was performed using summary data from the GWAS database. We conducted a two-sample Mendelian randomization (MR) study of these two diseases. Inverse variance weighting (IVW) was used as the primary analysis method, with MR Egger, weighted median, and weighted mode used as supplementary methods. Sensitivity analyses were performed using Cochran's Q test, MR-Egger intercept, MR-Pleiotropy RESidual Sum and outlier methods, leave-one-out analysis, and funnel plots. Results: A total of 11 single nucleotide polymorphisms (SNPs)were identified for instrumental variables(IVs) for MR analysis.IVW found that T1DM was causally associated with AS ((IVW: OR = 1.0006 (95% CI 1.0001, 1.0011), p = 0.0057; MR-Egger: OR = 1.0003 (95% CI 0.9995, 1.0012), p = 0.4147; weighted median: OR = 1.0006 (95% CI 1.0003, 1.0008), p = 0.0001; weighted mode: OR = 1.0007 (95% CI 1.0005, 1.0009), p = 0.0001). No horizontal pleiotropy was found for the MR-Egger intercept, and leave -one-out analysis found that the results remained stable after the removal of individual SNPs. Conclusion: The results of the two-sample MR analysis supported a causal relationship between T1DM and AS risk.
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Enfermedades Autoinmunes , Diabetes Mellitus Tipo 1 , Espondilitis Anquilosante , Humanos , Diabetes Mellitus Tipo 1/genética , Espondilitis Anquilosante/epidemiología , Espondilitis Anquilosante/genética , Estudio de Asociación del Genoma Completo , Análisis de la Aleatorización Mendeliana , NonoxinolRESUMEN
Postoperative abdominal adhesion is a very common and serious complication, resulting in pain, intestinal obstruction and heavy economic burden. Post-injury inflammation that could activate the coagulation cascade and deposition of fibrin is a major cause of adhesion. Many physical barrier membranes are used to prevent abdominal adhesion, but their efficiency is limited due to the lack of anti-inflammatory activity. Here, an electrospinning membrane composed of poly(lactic-co-glycolic acid) (PLGA) providing support and mechanical strength and chondroitin sulfate (CS) conferring anti-inflammation activity is fabricated for preventing abdominal adhesion after injury. The PLGA/CS membrane shows a highly dense fiber network structure with improved hydrophilicity and good cytocompatibility. Importantly, the PLGA/CS membrane with a mass ratio of CS at 20% provides superior anti-adhesion efficiency over a native PLGA membrane and commercial poly(D, L-lactide) (PDLLA) film in abdominal adhesion trauma rat models. The mechanism is that the PLGA/CS membrane could alleviate the local inflammatory response as indicated by the promoted percentage of anti-inflammatory M2-type macrophages and decreased expression of pro-inflammatory factors, such as IL-1ß, TNF-α and IL-6, resulting in the suppression of the coagulation system and the activation of the fibrinolytic system. Furthermore, the deposition of fibrin at the abdominal wall was inhibited, and the damaged abdominal tissue was repaired with the treatment of the PLGA/CS membrane. Collectively, the PLGA/CS electrospinning membrane is a promising drug-/cytokine-free anti-inflammatory barrier for post-surgery abdominal adhesion prevention and a bioactive composite for tissue regeneration.
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Sulfatos de Condroitina , Glicoles , Humanos , Ratas , Animales , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Adherencias Tisulares/prevención & control , Adherencias Tisulares/metabolismo , Antiinflamatorios/farmacologíaRESUMEN
BACKGROUND: Chronic inflammation and lack of angiogenesis are the important pathological mechanisms in Deep Tissue Injury (DTI). Curcumin is a well-known anti-inflammatory and antioxidant agent. However, curcumin is unstable under acidic and alkaline conditions and can be rapidly metabolized and excreted in the bile, which shortens its bioactivity and efficacy. OBJECTIVE: This study aimed to prepare curcumin-loaded poly (lactic-co-glycolic acid) nanoparticles (CPNPs) and to elucidate the protective effects and underlying mechanisms of wound healing in DTI models. METHODS: CPNPs were evaluated for particle size, biocompatibility, in vitro drug release and their effect on in vivo wound healing. RESULTS: The results of in vivo wound closure analysis revealed that CPNP treatments significantly improved wound contraction rates (p<0.01) at a faster rate than the other three treatment groups. H&E staining revealed that CPNP treatments resulted in complete epithelialization and thick granulation tissue formation. In contrast, control groups resulted in a lack of compact epithelialization and persistence of inflammatory cells within the wound sites. Quantitative real-time PCR analysis showed that treatment with CPNPs suppressed IL-6 and TNF-α mRNA expression, and up-regulated TGF-ß, VEGF-A and IL-10 mRNA expression. Western blot analysis showed up-regulated protein expression of TGF-ß, VEGF-A and phosphorylated-STAT3. CONCLUSION: Our results showed that CPNPs enhanced wound healing in DTI models through modulation of the JAK2/STAT3 signalling pathway and subsequent upregulation of pro-healing factors.
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Curcumina , Nanopartículas , Animales , Liberación de Fármacos , Ratones , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Cicatrización de HeridasRESUMEN
Cephalometric methods have been used to evaluate morphometric measurements of the mandible and quantify sex-related anatomical features; however, most studies to date employ a limited set of location-specific measurements without considering the entire three-dimensional anatomy of the mandible. The aims of this study were to develop statistical shape models (SSMs) of partially edentulous male and female mandibles to evaluate inter-subject morphological variability and secondly to assess the effectiveness of discrete clinical morphometric measurements in prediction of complete three-dimensional mandible geometry. Computed tomography images of forty partially edentulous female and twenty-five male subjects were obtained, and SSM developed using mesh fitting, rigid body registration and principal component analysis. Analysis of female and male SSMs showed that the variation along their first principal components was size-related. Sex-differentiating pure shape variations were found along the first principal component of size-normalised SSM and were observed to be most prominent in the symphysis and posterior ramus regions of the mandible. Seven morphometric measurements were found to characterise female and male shape prediction optimally. The capability to rapidly generate accurate patient-specific shape-predictive models of the mandible may be useful for implant development and pre-operative planning, particularly in the absence of bony structures following trauma or tumour resection.
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Simulación por Computador , Mandíbula/anatomía & histología , Adulto , Puntos Anatómicos de Referencia , Teorema de Bayes , Cefalometría , Femenino , Humanos , Masculino , Persona de Mediana Edad , Modelos Anatómicos , Modelos Estadísticos , Análisis de Componente Principal , Caracteres SexualesRESUMEN
Purpose The primary purpose of this study was to explore the audiovisual speech perception strategies.80.23.47 adopted by normal-hearing and deaf people in processing familiar and unfamiliar languages. Our primary hypothesis was that they would adopt different perception strategies due to different sensory experiences at an early age, limitations of the physical device, and the developmental gap of language, and others. Method Thirty normal-hearing adults and 33 prelingually deaf adults participated in the study. They were asked to perform judgment and listening tasks while watching videos of a Uygur-Mandarin bilingual speaker in a familiar language (Standard Chinese) or an unfamiliar language (Modern Uygur) while their eye movements were recorded by eye-tracking technology. Results Task had a slight influence on the distribution of selective attention, whereas subject and language had significant influences. To be specific, the normal-hearing and the d10eaf participants mainly gazed at the speaker's eyes and mouth, respectively, in the experiment; moreover, while the normal-hearing participants had to stare longer at the speaker's mouth when they confronted with the unfamiliar language Modern Uygur, the deaf participant did not change their attention allocation pattern when perceiving the two languages. Conclusions Normal-hearing and deaf adults adopt different audiovisual speech perception strategies: Normal-hearing adults mainly look at the eyes, and deaf adults mainly look at the mouth. Additionally, language and task can also modulate the speech perception strategy.
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Sordera , Percepción del Habla , Adulto , Tecnología de Seguimiento Ocular , Audición , Humanos , Lenguaje , BocaRESUMEN
BACKGROUND AND PURPOSE: New capillaries are essential for deep tissue pressure injury wound healing. Tazarotene is a recently discovered small molecule drug and functions to promote neovascularization and tissue repair. At present, the application of tazarotene in the repair of pressure injuries has not previously been investigated. This study used poly (lactic-co-glycolic acid) (PLGA) as nanoparticle carriers loaded with tazarotene (Ta/PLGA NPs) for drug delivery and to overcome shortcomings associated with the low water solubility, short half-life, easy photolysis and low bioavailability of tazarotene itself. METHODS: The physicochemical properties, drug release and bioactivity of Ta/PLGA NPs were examined in vitro by transmission electron microscope, spectrophotometry and cell assays. Mouse models of deep tissue pressure injuries (DTPI) were established and the therapeutic effects and mechanisms of Ta/PLGA NPs in local wound repair were studied. RESULTS: The results showed that Ta/PLGA NPs were of uniform size and distribution and were non-toxic both in vitro and in vivo. In vivo experiments suggested that Ta/PLGA NPs significantly promoted DTPI wound repair through activation of the VEGF/VEGFR-Notch1/DLL4 signaling pathway. CONCLUSION: This study highlights the potential clinical significance of implementation of tazarotene small molecule drugs in combination with effective biomaterial carriers for the treatment of chronic refractory wounds, such as DTPI.
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Nanopartículas , Ácido Poliglicólico , Úlcera por Presión , Animales , Ratones , Portadores de Fármacos , Ácido Láctico , Ácidos Nicotínicos , Tamaño de la Partícula , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Transducción de Señal , Factor A de Crecimiento Endotelial Vascular , Úlcera por Presión/tratamiento farmacológicoRESUMEN
Polycation carriers hold great potential in gene therapy. However, they usually suffer from obvious cytotoxicity and unsatisfactory transfection efficiency. In this report, a series of fluorobenzene substituted and thioacetal contained polycations (TAEA-S-xF) were prepared to explore novel alternatives for safe and efficient non-viral polymeric gene vectors. The reactive oxygen species (ROS)-responsive property of thioacetal moieties together with the fluorine effect were hope to bring the vector better performance in gene delivery process. These materials could efficiently condense DNA into nanoparticles with proper size and surface potential. The structure-activity relationship of these materials was systematically investigated, and the In vitro transfection results revealed that the amount of fluorine atoms on the linkage plays important role to ensure the transfection efficiency and serum tolerance. The ROS-responsive behavior was verified by NMR, gel electrophoresis experiment and dynamic light scattering (DLS) assay. Cytotoxicity assay results also suggest that these ROS-degradable polycations show good biocompatibility in response to higher ROS level in cancer cells. Among these fluorinated polymers, the one with the most fluorine atoms showed the best transfection efficiency, which was up to 54 times higher than polyethyleneimine (PEI) 25â¯kDa. Mechanism studies reveal that its better performance may come from good cellular uptake and endosome escape ability.
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Técnicas de Transferencia de Gen , Vectores Genéticos/química , Poliaminas/química , Polietileneimina/química , Especies Reactivas de Oxígeno/metabolismo , Animales , Células CHO , Cricetulus , Relación Dosis-Respuesta a Droga , Flúor/química , Flúor/metabolismo , Vectores Genéticos/síntesis química , Vectores Genéticos/metabolismo , Halogenación , Humanos , Estructura Molecular , Células PC-3 , Poliaminas/síntesis química , Poliaminas/metabolismo , Polielectrolitos , Polietileneimina/síntesis química , Polietileneimina/metabolismo , Relación Estructura-ActividadRESUMEN
Fluorinated biomaterials have been reported to have promising features as non-viral gene carriers. In this study, a series of fluorinated polymeric gene carriers were synthesized via Michael addition from low molecular weight polyethyleneimine (PEI) and fluorobenzoic acids (FBAs)-based linking compounds with different numbers of fluorine atoms. The structure-activity relationship (SAR) of these materials was systematically investigated. SAR studies showed that fluorine could screen the positive charge of these polymers. However, this shielding effect of fluorine would endow fluorinated polymers with good balance between DNA condensation and release. In vitro transfection results suggested that these fluorinated polymers could mediate efficient gene delivery. Flow cytometry and confocal microscopy studies demonstrated that more efficient cell uptake could be achieved by fluorinated materials with more fluorine atoms. Cytotoxicity assays showed that these fluorinated materials exhibited very low cytotoxicity even at high mass ratios. This study demonstrates that FBA-based fluorinated biopolymers have the potential for practical application.
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Polímeros de Fluorocarbono/uso terapéutico , Polietileneimina , Transfección/métodos , Línea Celular , Células/metabolismo , ADN/metabolismo , ADN/farmacocinética , Citometría de Flujo , Polímeros de Fluorocarbono/toxicidad , Terapia Genética/métodos , Humanos , Microscopía Confocal , Peso Molecular , Relación Estructura-ActividadRESUMEN
Non-rigid registration is a common part of bioengineering model-generation workflows. Compared to common mesh-based methods, radial basis functions can provide more flexible deformation fields due to their meshless nature. We introduce an implementation of RBF non-rigid registration with iterative knot-placement to adaptively reduce registration error. The implementation is validated on surface meshes of the femur, hemi-pelvis, mandible, and lumbar spine. Mean registration surface errors ranged from 0.37 to 0.99 mm, Hausdorff distance from 1.84 to 2.47 mm, and DICE coefficients from 0.97 to 0.99. The implementation is available for use in the free and open-source GIAS2 library.
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Algoritmos , Procesamiento de Imagen Asistido por Computador/métodos , Huesos/anatomía & histología , Humanos , Reproducibilidad de los ResultadosRESUMEN
A reductase-cleavable and thermo-responsive star-shaped polymer nanogel was prepared via an "arm-first" atom transfer radical polymerization approach. The nanogel consists of a thermo- and redox-sensitive core and a zwitterionic copolymer block. The dual sensitive core is composed of poly(N-isopropylacrylamide) that is formed by disulfide crosslinking of N-isopropylacrylamide. The zwitterionic copolymer block contains a poly(sulfobetaine methacrylate) component, a known anti-adsorptive moiety that extends blood circulation time, and a lactose motif of poly(2-lactobionamidoethyl methacrylamide) that specifically targets the asialoglycoprotein receptors (ASGP-Rs) of hepatoma. Doxorubicin (DOX) was encapsulated into the cross-linked nanogels via solvent extraction/evaporation method and dialysis; average diameter of both blank and DOX-loaded nanogels was ~120 nm. The multi-responsiveness of nanogel drug release in different temperatures and redox conditions was assessed. After 24 h, DOX release was only ~20% at 30°C with 0 mM glutathione (GSH), whereas over 90% DOX release was observed at 40°C and 10 mM GSH, evidence of dual responsiveness to temperature and reductase GSH. The IC50 value of DOX-loaded nanogels was much lower in human hepatoma (HepG2) cells compared to non-hepatic HeLa cells. Remarkably, DOX uptake of HepG2 cells differed substantially in the presence and absence of galactose (0.31 vs 1.42 µg/mL after 48 h of incubation). The difference was non-detectable in HeLa cells (1.21 vs 1.57 µg/mL after 48 h of incubation), indicating that the overexpression of ASGP-Rs leads to the DOX-loaded lactosylated nanogels actively targeting hepatoma. Our data indicate that the lactose-decorated star-shaped nanogels are dual responsive and hepatoma targeted, and could be employed as hepatoma-specific anti-cancer drug delivery vehicle for cancer chemotherapy.
Asunto(s)
Antibióticos Antineoplásicos/farmacocinética , Carcinoma Hepatocelular/tratamiento farmacológico , Sistemas de Liberación de Medicamentos/métodos , Neoplasias Hepáticas/tratamiento farmacológico , Polietilenglicoles/química , Polietileneimina/química , Resinas Acrílicas/síntesis química , Resinas Acrílicas/química , Antibióticos Antineoplásicos/administración & dosificación , Receptor de Asialoglicoproteína/metabolismo , Carcinoma Hepatocelular/metabolismo , Doxorrubicina/administración & dosificación , Doxorrubicina/farmacocinética , Liberación de Fármacos , Células HeLa/efectos de los fármacos , Células Hep G2/efectos de los fármacos , Humanos , Lactosa/análogos & derivados , Lactosa/química , Neoplasias Hepáticas/metabolismo , Espectroscopía de Resonancia Magnética , Microscopía Electrónica de Transmisión , Terapia Molecular Dirigida/métodos , Nanogeles , Oxidación-Reducción , Polietilenglicoles/administración & dosificación , Polietilenglicoles/farmacocinética , Polietileneimina/administración & dosificación , Polietileneimina/farmacocinética , Polimerizacion , Ácidos Polimetacrílicos/químicaRESUMEN
There is a need to identify and select new promising immunodominant antigens that have the ability to provide protective immunity against E. coli causing bovine mastitis. Recently we showed that f17a was found to be the most prevalent and crucial virulent factor among the pathogenic E. coli isolated from bovine mastitis. Here, in this report, the recombinant F17A based subunit vaccine adjuvant with MF59 was tested for immunogenicity against E. coli in a murine model. The vaccinated mice did not show any abnormal behavioral changes and histopathological lesions after vaccination. The specific antibody level against F17A was significantly higher in MF59-adjuvant-group, and also lasted for longer duration with a significant (P < 0.01) production level of IgG1 and IgG2a. Moreover, we noted higher survival rate in mice injected with F17A-MF59-adjuvant group after challenging with the clinical E. coli strain. Our findings of bacterial clearance test revealed that elimination rate from liver, spleen, and kidney in MF59-adjuvant-group was significantly higher than the control group. Finally, the proportion of CD4+T cells was increased, while CD8+ was decreased in MF59-adjuvant group. In conclusion, the current study reveals the capability of F17A-MF59 as a potential vaccine candidate against pathogenic E. coli causing mastitis in dairy animals.