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This paper presents a maskless lithography system that can perform three-dimensional (3D) ultraviolet (UV) patterning on a photoresist (PR) layer. After PR developing processes, patterned 3D PR microstructures over a large area are obtained. This maskless lithography system utilizes an UV light source, a digital micromirror device (DMD), and an image projection lens to project a digital UV image on the PR layer. The projected UV image is then mechanically scanned over the PR layer. An UV patterning scheme based on the idea of obliquely scanning and step strobe lighting (OS3L) is developed to precisely control the spatial distribution of projected UV dose, such that desired 3D PR microstructures can be obtained after PR development. Two types of concave microstructures with truncated conical and nuzzle-shaped cross-sectional profiles are experimentally obtained over a patterning area of 160 ×115 mm2. These patterned microstructures are then used for replicating nickel molds and for mass-production of light-guiding plates used in back-lighting and display industry. Potential improvements and advancements of the proposed 3D maskless lithography technique for future applications will be addressed.
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Tumor-associated macrophages (TAMs) in the tumor microenvironment have been associated with enhanced tumor progression. In this study, we investigated the role and molecular mechanisms of MALAT1 in TAMs derived from thyroid cancer. The expression of MALAT1 and FGF2 in thyroid cancer tissues and cells were measured by quantitative real-time PCR and Western blot. TAMs were transfected with indicated constructs. Then the culture medium (CM) from TAMs was harvested for assay. Secreted FGF2 protein levels and TNF-α, IL-12, and IL-10 levels were detected by ELISA. The cell proliferation, migration, and invasion of FTC133 cells were determined with a CCK-8 assay and a Transwell assay, respectively. In addition, HUVEC vasculature formation was measured by matrigel angiogenesis assay. The higher levels of MALAT-1 and FGF2 were observed in thyroid cancer tissues and in thyroid cancer cells compared to that in the control. Besides, in the presence of si-MALAT1, the levels of TNF-α and IL-12 were significantly up-regulated whereas IL-10 was down-regulated in the CM from TAMs. Moreover, down-regulation of MALAT1 in TAMs reduced proliferation, migration, and invasion of FTC133 cells and inhibited angiogenesis. However, overexpression of FGF2 blocked the effects of MALAT1 siRNAs on cell migration, invasion, and angiogenesis. Our results suggest that MALAT1-mediated FGF2 protein secretion from TAMs inhibits inflammatory cytokines release, promotes proliferation, migration, and invasion of FTC133 cells and induces vasculature formation. J. Cell. Biochem. 118: 4821-4830, 2017. © 2017 Wiley Periodicals, Inc.
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Factor 2 de Crecimiento de Fibroblastos/metabolismo , Macrófagos/metabolismo , Proteínas de Neoplasias/metabolismo , Neovascularización Patológica/metabolismo , ARN Largo no Codificante/metabolismo , ARN Neoplásico/metabolismo , Neoplasias de la Tiroides/metabolismo , Anciano , Línea Celular Tumoral , Femenino , Factor 2 de Crecimiento de Fibroblastos/genética , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Células Endoteliales de la Vena Umbilical Humana/patología , Humanos , Macrófagos/patología , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Proteínas de Neoplasias/genética , Neovascularización Patológica/genética , Neovascularización Patológica/patología , ARN Largo no Codificante/genética , ARN Neoplásico/genética , Neoplasias de la Tiroides/irrigación sanguínea , Neoplasias de la Tiroides/genética , Neoplasias de la Tiroides/patologíaRESUMEN
As a means of visual function testing and visual related mutants screening, the optokinetic response (OKR) and the optomotor response (OMR) behaviour tests are simple and effective tools for visual functional testing, which have been widely used in studying zebrafish larvae. However, adult zebrafish OKR analysis method is rarely reported. In this study, the methods of inducing adult zebrafish OKR behaviour, as well as tracking the movement of eyes using Pattern Match approaches, are presented. The quantitative measurement of the adult zebrafish OKR behaviour was successfully established. Using these methods, the binocular vision area was found to make a certain contribution to OKR behaviour. Moreover, the monocular vision of adult zebrafish showed a certain degree of directional sensitivity to moving gratings. Such approaches can also be applied to the zebrafish larvae OKR. The abnormal OKR behaviour phenomenon of period1b mutant larvae fish was detected.
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Nistagmo Optoquinético/fisiología , Pez Cebra/fisiología , Animales , Conducta Animal/fisiologíaRESUMEN
Under the background of rapid urbanization, the problem of fragmented habitat patches in economically developed areas is particularly prominent, where biodiversity is seriously threa-tened. Therefore, the construction of ecological network is an important measure to connect habitat patches and protect biological habitats. We extracted ecological source areas of Foshan City by using the connectivity index and morphological spatial pattern analysis (MSPA). Potential ecological corridors were identified based on InVEST model and minimal cumulative resistance (MCR). Combining the radiation channels extracted from hydrological analysis to build an ecological network in Foshan City. The ecological network was optimized by adding ecological source areas, stepping stones, and identifying fracture points. Finally, the network before and after optimization was evalua-ted from the aspects of both structure and function based on network analysis method and circuit theory. The results showed that ecological network in Foshan City was composed of 10 ecological source areas, 8 important corridors, 37 general corridors, and 11 radiation channels. After optimization, 7 new ecological source areas, 17 planning corridors, 13 stepping stones, and 80 fracture points were added. After optimization, the ecological network closure, the line rate index and the connection degree index were 0.59, 1.94, and 0.73, respectively. The maximum current density increased from 1.39 to 9.66 after optimization, indicating that the optimized ecological network structure was more perfect and highly connective.
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Conservación de los Recursos Naturales , Ecosistema , Biodiversidad , China , CiudadesRESUMEN
Nitrate (NO3-) reduction reaction (NtRR) is considered as a green alternative method for the conventional method of NH3 synthesis (Haber-Bosch process), which is known as a high energy consuming and large CO2 emitting process. Herein, the copper nanodendrites (Cu NDs) grown along with the {200} facet as an efficient NtRR catalyst have been successfully fabricated and investigated. It exhibited high Faradaic efficiency of 97% at low potential (-0.3 V vs RHE). Furthermore, the 15NO3- isotope labeling method was utilized to confirm the formation of NH3. Both experimental and theoretical studies showed that NtRR on the Cu metal nanostructure is a facet dependent process. Dissociation of NO bonding is supposed to be the rate-determining step as NtRR is a spontaneously reductive and protonation process for all the different facets of Cu. Density functional theory (DFT) calculations revealed that Cu{200} and Cu{220} offer lower activation energy for dissociation of NO compared to that of Cu{111}.
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This paper proposes a five degrees-of-freedom measurement system for measuring geometric errors of the rotary axis. To align the measured rotary axis with the reference axis, a diode laser is used to represent the rotary axis of the measured rotation stage. Based on the proposed measurement system, a model for separating the position independent geometric errors and position dependent geometric errors of the measured rotary axis from the measured value is established and verified by measurement experiments. The results of measurement experiments repeated for five times show that the measurement uncertainty of the proposed measurement system is less than ±1.6 µm for radial motion, the measurement uncertainty is less than ±1.7 arc sec for tilt motion, and the measurement uncertainty is less than ±1.3 arc sec for angle position.
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Several wearable devices have already been commercialized and are likely to open up a new life pattern for consumers. However, the limited energy capacity and lifetime have made batteries the bottleneck in wearable technology. Thus, there have been growing efforts in the area of self-powered wearables that harvest ambient mechanical energy directly from surroundings. Herein, we demonstrate a woven triboelectric nanogenerator (WTENG) utilizing electrospun Polyvinylidene fluoride (PVDF) nanofibers and commercial nylon cloth to effectively harvest mechanical energy from human motion. The PVDF nanofibers were fabricated using a highly scalable multi-nozzle far-field centrifugal electrospinning protocol. We have also doped the PVDF nanofibers with small amounts of multi-walled carbon nanotubes (MWCNT) to improve their triboelectric performance by facilitating the growth of crystalline ß-phase with a high net dipole moment that results in enhanced surface charge density during contact electrification. The electrical output of the WTENG was characterized under a range of applied forces and frequencies. The WTENG can be triggered by various free-standing triboelectric layers and reaches a high output voltage and current of about 14 V and 0.7 µA, respectively, for the size dimensions 6 × 6 cm. To demonstrate the potential applications and feasibility for harvesting energy from human motion, we have integrated the WTENG into human clothing and as a floor mat (or potential energy generating shoe). The proposed triboelectric nanogenerator (TENG) shows promise for a range of power generation applications and self-powered wearable devices.
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HYPOTHESIS: Superparamagnetic iron oxide nanoparticles (SPIONs) are extensively used as building block of colloidal nanocomposites for biomedical applications. Strategies employed to embed them in a biodegradable and biocompatible polymer matrix often fail to achieve a high density of loading which would greatly benefit to applications such as imaging and hyperthermia. In this study, poly(acrylic acid) coated SPION (γ-Fe2O3-PAA) are self-assembled with hydrolysable poly(serine ester) by electrostatic complexation, leading to perfectly defined spherical particles with ultra-high density of magnetic material and an ability to auto-degrade into individual SPION and biocompatible byproducts. EXPERIMENTS: Self-assembly and auto-degradation of γ-Fe2O3-PAA/poly(serine ester) and γ-Fe2O3-PAA/poly(serine ester)-b-PEG colloidal particles are studied by light scattering and microscopy. Colloidal stability in bio-fluids, hyperthermia under alternating magnetic field, cellular uptake, cytotoxicity and degradation of γ-Fe2O3-PAA/poly(serine ester)-b-PEG in living cells are investigated. FINDINGS: A remarkably slow electrostatic complexation leads to dense superparamagnetic γ-Fe2O3-PAA/poly(serine ester)-b-PEG polyion complexes (PICs) with controlled sizes (150-500â¯nm) and times of degradation in aqueous solvents (700-5000â¯h). The material shows good sustainability during hyperthermia, is well taken up by MC3T3 cells and non-cytotoxic. TEM images reveal a mechanism of degradation by "peeling" and fragmentation. In cells, PICs are reduced into individual SPIONs within 72â¯h.
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Materiales Biocompatibles/química , Coloides/química , Fenómenos Magnéticos , Nanopartículas de Magnetita/química , Péptidos/química , Polímeros/química , Resinas Acrílicas/síntesis química , Resinas Acrílicas/química , Animales , Dispersión Dinámica de Luz , Células Hep G2 , Humanos , Hipertermia Inducida , Ratones , Péptidos/síntesis química , Polietilenglicoles/síntesis química , Polietilenglicoles/química , Polímeros/síntesis química , Pruebas de ToxicidadRESUMEN
Under a controlled adsorption environment, L-cysteine molecules can be chemically adsorbed to the dendritic silver (Ag-D) surface by electrochemical methods with different functional groups. It is verified by surface-enhanced Raman spectroscopy that under alkaline conditions (pH = 13.50), the two functional groups of thiol and acid are simultaneously adsorbed on the surface of Ag-D, while NH2 is far from the surface; under acidic conditions (pH = 1.67), adsorption behavior suggests that both NH3+ and COO- are oriented toward the Ag-D surface, and that SH is far from the surface. The structure of L-cysteine adsorption under acidic conditions can be further verified by the addition of an L-cysteine molecule through light-induced coupling reaction to form cystine. Finally, in-situ two-dimensional Raman scattering spectroscopy confirmed the feasibility and uniformity of the coupling reaction.
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OBJECTIVE: To discuss the feasibility of biliary reconstruction and rehabilitation after transection injury of biliary duct by laparoscopy. METHODS: The clinical data of 24 cases receiving biliary reconstruction after transection injury of biliary duct by laparoscopy were analyzed retrospectively from August 2002 to April 2008, including operation indications, contraindications, related operation skills and so on. In these 24 cases, the reasons of transection of biliary duct as followed: 15 cases were pancreaticoduodenectomy, 6 cases were resection of the choledochal cyst, 1 case was resection of high cholangiocarcinoma, 1 case was cholecystectomy and 1 case was resection of gastric cancer. RESULTS: Biliary reconstruction and rehabilitation was successfully completed in 24 cases by laparoscopy. There was 1 case of bile leakage and no duct stenosis complications. CONCLUSIONS: Biliary reconstruction and rehabilitation by laparoscopy was feasible and safe procedure, has a high successful rate, and deserves further clinical trials in hospitals.
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Anastomosis Quirúrgica/métodos , Conductos Biliares/lesiones , Conductos Biliares/cirugía , Laparoscopía , Adolescente , Adulto , Anciano , Niño , Preescolar , Estudios de Factibilidad , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
OBJECTIVE: To explore the feasibility and the effect of laparoscopic hepatectomy for primary liver cancer(PLC). METHODS: A retrospective study on 61 cases of laparoscopic hepatectomy for PLC was made between November 2002 and June 2007, among which there were 49 male and 12 female, aged from 14 to 71 years. All patients were diagnosed as PLC by type-B ultrasonic, CT or MRI, and APF. RESULTS: Fifty-six patients were completed laparoscopically successfully. Five cases underwent conversion to open operation because of hemorrhage. The mean operative time was 60 min (30-150 min). The mean blood loss was 450 ml (100-2000 ml). The mean hepatic portal block time was 20 min (15-30 min). All the patients had excellent recovery without any postoperative surgical complications. The patients were mobilized out of the bed in 24 hours. Oral intake of food started in 1 to 3 days. The average postoperative hospital stay was 6.6 d (5-10 d). CONCLUSION: Laparoscopic hepatectomy for PLC is safe and feasible by using hepatic portal block instrument.
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Hepatectomía/métodos , Laparoscopía , Neoplasias Hepáticas/cirugía , Adolescente , Adulto , Anciano , Estudios de Factibilidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
The circadian clock is an endogenous and entrainable time-keeping mechanism with a period of approximately 24 h, operated by transcription/translation feedback loops composed of circadian clock genes and their proteins. The visual system displays robust circadian changes. Relatively little, however, is known about the mechanisms underlying visual circadian rhythmicity. Zebrafish period1b (per1b), as a canonical circadian clock gene, is involved in circadian regulation. Here, we observed that zebrafish per1b mutants exhibit visual defects including reduced behavioral contrast sensitivity and significant retinal dopaminergic deficiency. Further, partially damaged dopaminergic interplexiform cells in wild-type larvae also led to reduced behavioral contrast sensitivity, while exogenous dopamine administration effectively restored the contrast sensitivity of per1b mutants. Taken together, these results suggest that retinal dopaminergic deficiency derived from loss of per1b results in visual defects in zebrafish. ABBREVIATIONS: per1b, period1b; per, period; per1, period1; per2, period2; per3, period3; ERG, electroretinogram; DA-IPCs, dopaminergic interplexiform cells; IRBP, interphotoreceptor retinoid binding protein; MS-222, methane-sulfonate; USTC, University of Science and Technology of China; OKR, optokinetic response; dpf, day postfertilization; 6-OHDA, 6-hydroxydopamine; TH, tyrosine hydroxylase; DA, dopaminergic; INL, inner nuclear; IPL, innerplexiform layers; hpf, hours postfertilization; cpd, cycle per degree; ADHD, attention deficit and hyperactivity disorder.
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Proteínas CLOCK/metabolismo , Relojes Circadianos/fisiología , Ritmo Circadiano/fisiología , Proteínas Circadianas Period/metabolismo , Proteínas de Pez Cebra/genética , Animales , Proteínas CLOCK/genética , Dopamina/metabolismo , Proteínas Circadianas Period/genética , Retina/metabolismo , Tirosina 3-Monooxigenasa/metabolismo , Pez Cebra/genéticaRESUMEN
Zebrafish is an excellent model to study central nervous system (CNS) axonal degeneration and regeneration since we can observe these processes in vivo and in real time in transparent larvae. Previous studies have shown that Mauthner cell (M-cell) axon regenerates poorly after mechanical spinal cord injury. Inconsistent with this result, however, we have found that M-cell possesses a great capacity for axon regeneration after two-photon laser ablation. By using ZEISS LSM 710 two-photon microscope, we performed specific unilateral axotomy of GFP labeled M-cells in the Tol-056 enhancer trap line larvae. Our results showed that distal axons almost degenerated completely at 24h after laser axotomy. After that, the proximal axons initiated a robust regeneration and many of the M-cell axons almost regenerated fully at 4days post axotomy. Furthermore, we also visualized that regenerated axons were remyelinated when we severed fluorescent dye labeled M-cells in the Tg (mbp:EGFP-CAAX) line larvae. Moreover, by single M-cell co-electroporation with Syp:EGFP and DsRed2 plasmids we observed synapses re-establishment in vivo during laser injury-induced axon re-extension which suggested re-innervation of denervated pathways. In addition, we further demonstrated that nocodazole administration could completely abolish this regeneration capacity. These results together suggested that in vivo time-lapse imaging of M-cell axon laser injury may provide a powerful analytical model for understanding the underlying cellular and molecular mechanisms of the CNS axon regeneration.
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Axones/fisiología , Microscopía Intravital/métodos , Larva/fisiología , Regeneración Nerviosa/fisiología , Remielinización/fisiología , Sinapsis/fisiología , Animales , Animales Modificados Genéticamente , Axotomía/efectos adversos , Imagen de Lapso de Tiempo/métodos , Pez CebraRESUMEN
BACKGROUND: Endoscopic thyroidectomy for minimally invasive thyroid surgery has been widely applied in the past decade. The present study aimed to evaluate the effects of single-port access transaxillary totally endoscopic thyroidectomy on the postoperative outcomes and functional parameters, including quality of life and cosmetic result in patients with papillary thyroid carcinoma (PTC). PATIENTS AND METHODS: Seventy-five patients with PTC who underwent endoscopic thyroidectomy via a single-port access transaxillary approach were included (experimental group). A total of 123 patients with PTC who were subjected to conventional open total thyroidectomy served as the control group. The health-related quality of life and cosmetic and satisfaction outcomes were assessed postoperatively. RESULTS: The mean operation time was significantly increased in the experimental group. The physiological functions and social functions in the two groups were remarkably augmented after 6 months of surgery. However, there was no significant difference in the scores of speech and taste between the two groups at the indicated time of 1 month and 6 months. In addition, the scores for appearance, satisfaction with appearance, role-physical, bodily pain, and general health in the experimental group were better than those in the control group at 1 month and 6 months after surgery. CONCLUSION: The single-port access transaxillary totally endoscopic thyroidectomy is safe and feasible for the treatment of patients with PTC. The subjects who underwent this technique have a good perception of their general state of health and are likely to participate in social activities. It is worthy of being clinically used for patients with PTC.
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BACKGROUND: Increasing evidence indicated that metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) acted as a key regulator in the proliferation and invasion of several cancers. However, the function of MALAT1 in the development of thyroid cancer has not been experimentally established. METHODS: The expression of MALAT1 and IQGAP1 in thyroid cancer tissues and cells were detected by quantitative real-time PCR and western blot. The effects of MALAT1 and IQGAP1 on the cell proliferation and invasion of thyroid cancer cells were detected with a 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium 4 (MTT) assay and a Transwell assay, respectively. FTC-133 or SW1736 transfected with si-MALAT1 or pcDNA-MALAT1 were injected subcutaneously into 4-week-olds BALB/c mice to examine the impact of MALAT1 on the tumor development of thyroid cancer in vivo. RESULTS: In this study, we discovered the higher level of MALAT-1 and expression of IQGAP1 in thyroid cancer tissues and in thyroid cancer cells compared to that in the control. MTT and Transwell assay showed that the proliferation and invasion of FTC-133 cells with MALAT-1 knockdown were inhibited. Moreover, MALAT-1 could upregulate the expression of IQGAP1 in thyroid cancer cells. In addition, IQGAP1 knockdown reversed the decreasing cell proliferation and invasion of thyroid cancer induced by MALAT-1 overexpression. Finally, the study in vivo verified that MALAT-1 promoted the tumor growth of thyroid cancer. CONCLUSION: Our study indicated that MALAT1 promoted the proliferation and invasion of thyroid cancer cells via regulating the expression of IQGAP1.
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Regulación Neoplásica de la Expresión Génica , ARN Largo no Codificante/metabolismo , Neoplasias de la Tiroides/genética , Neoplasias de la Tiroides/patología , Proteínas Activadoras de ras GTPasa/genética , Línea Celular Tumoral , Proliferación Celular , Humanos , Invasividad Neoplásica , ARN Largo no Codificante/genética , Tejido Subcutáneo/patología , Regulación hacia Arriba , Ensayos Antitumor por Modelo de Xenoinjerto , Proteínas Activadoras de ras GTPasa/metabolismoRESUMEN
Dendritic spines undergo continuous remodeling during development of the nervous system. Their stability is essential for maintaining a functional neuronal circuit. Spine dynamics and stability of cortical excitatory pyramidal neurons have been explored extensively in mammalian animal models. However, little is known about spiny interneurons in non-mammalian vertebrate models. In the present study, neuronal morphology was visualized by single-cell electroporation. Spiny neurons were surveyed in the Xenopus tadpole brain and observed to be widely distributed in the olfactory bulb and telencephalon. DsRed- or PSD95-GFP-expressing spiny interneurons in the olfactory bulb were selected for in vivo time-lapse imaging. Dendritic protrusions were classified as filopodia, thin, stubby, or mushroom spines based on morphology. Dendritic spines on the interneurons were highly dynamic, especially the filopodia and thin spines. The stubby and mushroom spines were relatively more stable, although their stability significantly decreased with longer observation intervals. The 4 spine types exhibited diverse preferences during morphological transitions from one spine type to others. Sensory deprivation induced by severing the olfactory nerve to block the input of mitral/tufted cells had no significant effects on interneuron spine stability. Hence, a new model was established in Xenopus laevis tadpoles to explore dendritic spine dynamics in vivo.
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Espinas Dendríticas/fisiología , Interneuronas/fisiología , Plasticidad Neuronal/fisiología , Bulbo Olfatorio/fisiología , Animales , Femenino , Interneuronas/citología , Larva , Bulbo Olfatorio/citología , Imagen de Lapso de Tiempo , Xenopus laevisRESUMEN
Monomethyl poly(ethylene glycol) (mPEG)-modified bovine serum albumin (BSA) conjugates (BSA-mPEG) were obtained by the mild Cu(I)-mediated cycloaddition reaction of azided BSA (BSA-N(3) ) and alkyne-terminated mPEG. The structure and characteristics of BSA-mPEG conjugates were thoroughly investigated. There were about two PEG chains conjugated onto each BSA molecule as determined by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) analysis. The intrinsic nonspecific binding ability of BSA was used for adsorption and sustained release of both rifampicn and 5-fluorouracil (5-FU). The helical structures of BSA were preserved to a large extent after modification and drug adsorption on BSA was confirmed via circular dichroism spectroscopy. Drugs adsorbed onto the conjugated formulation to a lesser extent than on BSA due to mPEG modification. The in vitro release of both rifampicin and 5-FU, however, indicated that BSA-mPEG can function as a drug carrier. Overall, the click reaction provided a convenient tool for the pegylation of BSA. The biological activity of the BSA-mPEG conjugates, including the drug transportation capacity and biocompatibility, were largely retained.
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Portadores de Fármacos , Polietilenglicoles/química , Albúmina Sérica Bovina/química , Dicroismo Circular , Electroforesis en Gel de Poliacrilamida , Fluorouracilo/administración & dosificación , Espectroscopía de Resonancia Magnética , Rifampin/administración & dosificación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Optokinetic response (OKR) is a behavior that an animal vibrates its eyes to follow a rotating grating around it. It has been widely used to assess the visual functions of larval zebrafish. Nevertheless, the standard protocol for larval fish is not yet readily applicable in adult zebrafish. Here, we introduce how to measure the OKR of adult zebrafish with our simple custom-built apparatus using a new protocol which is established in our lab. Both our apparatus and step-by-step procedure of OKR in adult zebrafish are illustrated in this video. In addition, the measurements of the larval OKR, as well as the optomotor response (OMR) test of adult zebrafish, are also demonstrated in this video. This OKR assay of adult zebrafish in our experiment may last for up to 4 hours. Such OKR test applied in adult fish will benefit to visual function investigation more efficiently when the adult fish vision system is manipulated.
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Medidas del Movimiento Ocular/instrumentación , Nistagmo Optoquinético/fisiología , Pez Cebra/fisiología , AnimalesRESUMEN
Cell penetrating peptides (CPPs) are promising tools for transducing presynthesized therapeutic molecules which possess low membrane permeability. The poor efficiency of cellular uptake and unexpected cellular localization are still the main obstacles to the development of drug delivery by using CPPs. In this study, we investigated the effect of a penetration enhancer, dimethylsulfoxide (DMSO), on the penetrating efficiency of a synthetic TAT peptide or the TAT fusion protein. FITC-labeled TAT and TAT-GFP were added to 10% DMSO or 100 microM chloroquine pretreated cells, fluorescence uptake into culturing cells was observed using fluorescence microscopy, FACS or quantitatively analyzed by a fluorescence spectrum. 10% DMSO treatment markedly increased internalization of TAT into cells and appeared in a well-distributed pattern throughout the cytosol and nucleus without membrane perforating or detectable cytotoxicity, the enhancement effect by 10% DMSO was reduced by endocytosis inhibitors including ammonium chloride and sodium azide. 10% DMSO also enhanced TAT-Apoptin induced apoptosis of carcinoma cells. These findings implicated that DMSO can be a novel delivery enhancer appropriate for CPP penetration.