RESUMEN
The bleomycin-induced scleroderma model is a well-established and dependable method for creating a mouse model of SSc (systemic sclerosis). In the field of skin connective tissue diseases, increasing evidence from clinical and animal experiments suggests that TLRs (Toll-like receptors) play an important role in several diseases. This study aimed to determine the role of TLR7 (Toll-like receptor 7) and TLR9 (Toll-like receptor 9) in the mechanisms of immune abnormalities and fibrosis in SSc. This study used TLR7-KO mice (TLR7-knockout mice with a balb/c background) and TLR9-KO mice (TLR9-knockout mice with a balb/c background) as well as WT mice (wild-type balb/c mice). All three kinds of mice were induced by BLM (bleomycin) in a scleroderma model as the experimental group; meanwhile, WT mice treated with PBS (phosphate-buffered saline) were used as the control group. We analyzed the fibrotic phenotype and the immunological abnormality phenotype of TLR7-deficient and TLR9-deficient mice in the SSc disease model using flow cytometry, RT-PCR (reverse transcription-polymerase chain reaction), a histological examination, and IHC (immunohistochemical staining). In a mouse model of SSc disease, the deletion of TLR7 attenuated skin and lung fibrosis, while the deletion of TLR9 exacerbated skin and lung fibrosis. The deletion of TLR7 resulted in a relative decrease in the infiltration and expression of various pro-inflammatory and fibrotic cells and cytokines in the skin. On the other hand, the deletion of TLR9 resulted in a relative increase in the infiltration and expression of various pro-inflammatory and cytokine-inhibiting cells and cytokines in the skin. Under the influence of pDCs (plasmacytoid dendritic cells), the balances of Beff/Breg (IL-6 + CD19 + B cell/IL-10 + CD19 + B cell), Th17/Treg (IL-17A + CD4 + T cell/Foxp3 + CD25 + CD4 + T cell), M1/M2 (CD86 + macrophage/CD206 + macrophage), and Th1/Th2 (TNFα + CD3 + CD4 + T cell/IL-4 + CD3 + CD4 + T cell) were biased towards the suppression of inflammation and fibrosis as a result of the TLR7 deletion. Comparatively, the balance was biased towards promoting inflammation and fibrosis due to the TLR9 deletion. In the SSc model, TLR7 promoted inflammation and fibrosis progression, while TLR9 played a protective role. These results suggest that TLR7 and TLR9 play opposite roles in triggering SSc to produce immune system abnormalities and skin fibrosis.
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Modelos Animales de Enfermedad , Ratones Noqueados , Esclerodermia Sistémica , Receptor Toll-Like 7 , Receptor Toll-Like 9 , Animales , Receptor Toll-Like 7/metabolismo , Receptor Toll-Like 7/genética , Esclerodermia Sistémica/metabolismo , Esclerodermia Sistémica/patología , Esclerodermia Sistémica/inmunología , Esclerodermia Sistémica/genética , Receptor Toll-Like 9/metabolismo , Receptor Toll-Like 9/genética , Ratones , Bleomicina/efectos adversos , Ratones Endogámicos BALB C , Citocinas/metabolismo , Piel/patología , Piel/metabolismo , Piel/inmunología , Fibrosis , Fibrosis Pulmonar/metabolismo , Fibrosis Pulmonar/patología , Fibrosis Pulmonar/etiología , Glicoproteínas de MembranaRESUMEN
Anti-human upstream-binding factor (anti-hUBF) antibodies have been reported predominantly in patients with connective tissue diseases (CTDs); these have also been reported in patients without CTDs such as hepatocellular carcinoma. Because of the low frequency of expression and few case reports, there is no consensus on the clinical significance of these antibodies. Thus, we aimed to examine the clinical features of patients with anti-hUBF antibodies and analyzed 1042 patients with clinically suspected CTDs. The presence of anti-hUBF antibodies was screened using immunoprecipitation assays. Of the 1042 patients, 19 (1.82%) tested positive for anti-hUBF antibodies; among them, 10 (56%) were diagnosed with undifferentiated CTD (UCTD), six with systemic sclerosis (SSc) and three with other diseases. Five of the 10 patients with UCTD were referred to our hospital with suspected SSc. None of the five patients fulfilled the 2013 American College of Rheumatology/European League Against Rheumatism classification criteria, but three scored seven points, a relatively high score. Six anti-hUBF-positive patients with SSc had a significantly lower modified Rodnan skin score (mRSS) than that of anti-hUBF-negative patients with SSc (2 [0-2] vs 7 [0-49], p < 0.01). Compared with anti-topoisomerase I-positive patients, anti-hUBF-positive patients had a significantly lower mRSS (2 [0-2] vs 13 [0-42], p < 0.01) and lower incidence of scleroderma renal crisis (0 of 6 vs 8 of 184, p < 0.01). Compared with anti-centromere-positive patients, anti-hUBF-positive patients had a higher incidence of interstitial lung disease (ILD), but the difference was not statistically significant (4 of 6 vs 19 of 239). In conclusion, anti-hUBF antibodies were predominantly detected in patients with CTDs and UCTD. In patients with CTDs, SSc exhibited a high ratio, displaying a lower mRSS and higher incidence of ILD. In patients with UCTD, careful follow-up is recommended as they may develop CTDs in the future.
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Proteínas Adaptadoras Transductoras de Señales , Autoanticuerpos , Factores de Transcripción , Humanos , Masculino , Femenino , Estudios Retrospectivos , Persona de Mediana Edad , Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Adulto , Anciano , Enfermedades del Tejido Conjuntivo/inmunología , Enfermedades del Tejido Conjuntivo/diagnóstico , Esclerodermia Sistémica/inmunología , Esclerodermia Sistémica/sangre , Esclerodermia Sistémica/diagnóstico , Esclerodermia Sistémica/complicaciones , Índice de Severidad de la Enfermedad , Enfermedades Indiferenciadas del Tejido Conectivo/inmunología , Enfermedades Indiferenciadas del Tejido Conectivo/complicacionesRESUMEN
To summarize the current therapies for skin cancers, the Japanese Skin Cancer Society issued the first guidelines for skin cancers, including melanoma, squamous cell carcinoma, basal cell carcinoma (BCC), and extramammary Paget's disease, in 2007. These guidelines were revised in 2015. Herein, we present the English version of the 2021 edition of the Japanese clinical guidelines for BCC. In the latest edition, all procedures were performed according to the Grading of Recommendations, Assessment, Development and Evaluation systems. The clinical questions that could not be answered were selected for further analysis. A comprehensive literature search, systematic review, and recommendations for each clinical question were determined by a multidisciplinary expert panel comprising dermatologists, a plastic and reconstructive surgeon, and a pathologist. Surgical resection is the gold-standard therapy of BCC. Radiotherapy or topical treatments, other than surgical resection, have been used in some cases. Patients with unresectable or metastatic BCC require systemic therapy. Novel agents, such as immune response modifiers or hedgehog pathway inhibitors, are emerging worldwide for the treatment of BCC. Based on these viewpoints, four relevant clinical questions regarding, surgical resection, radiotherapy, topical treatment, and systemic therapy, were raised in this report that aims to help clinicians select suitable therapies for their patients.
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Carcinoma Basocelular , Neoplasias Cutáneas , Humanos , Carcinoma Basocelular/terapia , Carcinoma Basocelular/patología , Carcinoma Basocelular/diagnóstico , Neoplasias Cutáneas/terapia , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/diagnóstico , Japón , Dermatología/normas , Sociedades Médicas/normas , Antineoplásicos/uso terapéutico , Pueblos del Este de AsiaRESUMEN
Interstitial lung disease (ILD) is recognized a prognostic factor and leading cause of death in patients with systemic sclerosis (SSc). The aim of the present study is to clarify factors at an initial visit that are associated with the deterioration of ILD in SSc patients with anti-topoisomerase I (anti-topo I) antibodies. This was a single-center, retrospective, observational study. Fifty-three consecutive SSc patients with anti-topo I antibodies were included in this study. Of the 53 patients, 43 had ILD at their initial visit, whereas 10 did not. We examined the clinical and immunological factors at an initial visit that were associated with the deterioration of ILD. The deterioration of ILD was defined as the administration of intravenous cyclophosphamide (IVCY) therapy. In this cohort, 45 (85%) patients had ILD at the time of the final observation, and only two who did not have ILD at their initial visit developed ILD during the follow-up period. Until the final observation, 26 (49%) patients received IVCY therapy for the progression of ILD. The age at onset, disease duration, SSc subtype, and skin score were similar between patients with and those without IVCY therapy. Approximately 60% (26 of 43) of patients with ILD at their initial visit received IVCY therapy. On the other hand, none of the 10 patients without ILD at their initial visit received IVCY therapy. Our multivariate analyses using Cox proportional hazards regression model revealed that the presence of ILD at an initial visit was an independent factor associated with the introduction of IVCY therapy (odds ratio, 2.8e+7 [95% confidence interval, 1.8e+17-uncalculated], p = 0.0048). Although anti-topo I antibodies are strongly associated with ILD, it was unlikely for SSc patients with anti-topo I antibodies to receive IVCY therapy when they did not have ILD at an initial visit.
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Enfermedades Pulmonares Intersticiales , Esclerodermia Sistémica , Humanos , Inmunosupresores , Estudios Retrospectivos , Japón/epidemiología , Resultado del Tratamiento , Enfermedades Pulmonares Intersticiales/diagnóstico , Enfermedades Pulmonares Intersticiales/tratamiento farmacológico , Enfermedades Pulmonares Intersticiales/etiología , Ciclofosfamida/uso terapéutico , Esclerodermia Sistémica/complicaciones , Esclerodermia Sistémica/tratamiento farmacológico , PulmónRESUMEN
Argonaute (AGO) proteins use small RNAs to recognize transcripts targeted for silencing in plants and animals. Many AGOs cleave target RNAs using an endoribonuclease activity termed 'slicing'. Slicing by DNA-guided prokaryotic AGOs has been studied in detail, but structural insights into RNA-guided slicing by eukaryotic AGOs are lacking. Here we present cryogenic electron microscopy structures of the Arabidopsis thaliana Argonaute10 (AtAgo10)-guide RNA complex with and without a target RNA representing a slicing substrate. The AtAgo10-guide-target complex adopts slicing-competent and slicing-incompetent conformations that are unlike known prokaryotic AGO structures. AtAgo10 slicing activity is licensed by docking target (t) nucleotides t9-t13 into a surface channel containing the AGO endoribonuclease active site. A ß-hairpin in the L1 domain secures the t9-t13 segment and coordinates t9-t13 docking with extended guide-target pairing. Results show that prokaryotic and eukaryotic AGOs use distinct mechanisms for achieving target slicing and provide insights into small interfering RNA potency.
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Arabidopsis , Proteínas Argonautas , Animales , Proteínas Argonautas/metabolismo , ARN Interferente Pequeño/genética , Interferencia de ARN , Arabidopsis/genética , Arabidopsis/metabolismo , Dominio Catalítico , ARN de Planta/genética , ARN de Planta/metabolismoRESUMEN
BACKGROUND: Pain is a complex experience with both sensory and affective dimensions, and the affective dimension can increase the risks of chronic pain development. It is thus critical to identify factors influencing the affective dimension of pain. OBJECTIVE: This study aimed to identify the relationship between the affective dimension of pain and disorder site (primary pain source). METHODS: Study participants were recruited from patients referred for physical therapy at an orthopedic outpatient clinic. Pain quality including the affective dimension, disorder site from descriptive medical diagnosis, pain intensity, duration from pain onset, and demographic data, was collected. A multivariable logistic regression model was constructed to analyze the relationship between the affective dimension of pain and the disorder site. RESULTS: A total of 282 participants were included. Cervical and lumbar spine disorders were significantly associated with an affective dimension of pain compared to limbs disorders when adjusted for age, sex, pain intensity, and duration from the onset in the regression model. CONCLUSIONS: Regardless of duration from the onset and other confounding factors, disorder site is correlated with the affective dimension of pain. Multidimensional pain assessment is crucial when clinicians evaluate patients with cervical and lumbar spine disorders, even in the acute phase.
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Dolor Crónico , Humanos , Estudios Transversales , Dimensión del DolorRESUMEN
Group IV materials are promising candidates for highly reliable and human-friendly thin-film thermoelectric generators, used for micro-energy harvesting. In this study, we investigated the synthesis and thermoelectric applications of a Ge-based ternary alloy thin film, Ge1-x-ySixSny. The solid-phase crystallization of the highly densified amorphous precursors allowed the formation of high-quality polycrystalline Ge1-x-ySixSny layers on an insulating substrate. The small compositions of Si and Sn in Ge1-x-ySixSny (x < 0.15 and y < 0.05) lowered the thermal conductivity (3.1 W m-1 K-1) owing to the alloy scattering of phonons, while maintaining a high carrier mobility (approximately 200 cm2 V-1 s-1). The solid-phase diffusion of Ga and P allowed us to control the carrier concentration to the order of 1019 cm-3 for holes and 1018 cm-3 for electrons. For both p- and n-type Ge1-x-ySixSny, the power factor peaked at x = 0.06 and y = 0.02, reaching 1160 µW m-1 K-2 for p-type and 2040 µW m-1 K-2 for n-type. The resulting dimensionless figure of merits (0.12 for p-type and 0.20 for n-type) are higher than those of most environmentally friendly thermoelectric thin films. These results indicate that group IV alloys are promising candidates for high-performance, reliable thin-film thermoelectric generators.
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The human SMC5/6 complex is a conserved guardian of genome stability and an emerging component of antiviral responses. These disparate functions likely require distinct mechanisms of SMC5/6 regulation. In yeast, Smc5/6 is regulated by its Nse5/6 subunits, but such regulatory subunits for human SMC5/6 are poorly defined. Here, we identify a novel SMC5/6 subunit called SIMC1 that contains SUMO interacting motifs (SIMs) and an Nse5-like domain. We isolated SIMC1 from the proteomic environment of SMC5/6 within polyomavirus large T antigen (LT)-induced subnuclear compartments. SIMC1 uses its SIMs and Nse5-like domain to localize SMC5/6 to polyomavirus replication centers (PyVRCs) at SUMO-rich PML nuclear bodies. SIMC1's Nse5-like domain binds to the putative Nse6 orthologue SLF2 to form an anti-parallel helical dimer resembling the yeast Nse5/6 structure. SIMC1-SLF2 structure-based mutagenesis defines a conserved surface region containing the N-terminus of SIMC1's helical domain that regulates SMC5/6 localization to PyVRCs. Furthermore, SLF1, which recruits SMC5/6 to DNA lesions via its BRCT and ARD motifs, binds SLF2 analogously to SIMC1 and forms a separate Nse5/6-like complex. Thus, two Nse5/6-like complexes with distinct recruitment domains control human SMC5/6 localization.
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Proteínas de Ciclo Celular , Proteínas Cromosómicas no Histona , Humanos , Proteínas de Ciclo Celular/metabolismo , Proteínas Cromosómicas no Histona/metabolismo , Proteómica , Compartimentos de Replicación ViralRESUMEN
The emergence of tyrosine kinase inhibitors as part of a front-line treatment has greatly improved the clinical outcome of the patients with Ph+ acute lymphoblastic leukemia (ALL). However, a portion of them still become refractory to the therapy mainly through acquiring mutations in the BCR-ABL1 gene, necessitating a novel strategy to treat tyrosine kinase inhibitor (TKI)-resistant Ph+ ALL cases. In this report, we show evidence that RUNX1 transcription factor stringently controls the expression of BCR-ABL1, which can strategically be targeted by our novel RUNX inhibitor, Chb-M'. Through a series of in vitro experiments, we identified that RUNX1 binds to the promoter of BCR and directly transactivates BCR-ABL1 expression in Ph+ ALL cell lines. These cells showed significantly reduced expression of BCR-ABL1 with suppressed proliferation upon RUNX1 knockdown. Moreover, treatment with Chb-M' consistently downregulated the expression of BCR-ABL1 in these cells and this drug was highly effective even in an imatinib-resistant Ph+ ALL cell line. In good agreement with these findings, forced expression of BCR-ABL1 in these cells conferred relative resistance to Chb-M'. In addition, in vivo experiments with the Ph+ ALL patient-derived xenograft cells showed similar results. In summary, targeting RUNX1 therapeutically in Ph+ ALL cells may lead to overcoming TKI resistance through the transcriptional regulation of BCR-ABL1. Chb-M' could be a novel drug for patients with TKI-resistant refractory Ph+ ALL.
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Subunidad alfa 2 del Factor de Unión al Sitio Principal/metabolismo , Proteínas de Fusión bcr-abl/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Animales , Antineoplásicos/farmacología , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/genética , Subunidad alfa 2 del Factor de Unión al Sitio Principal/antagonistas & inhibidores , Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Resistencia a Antineoplásicos/efectos de los fármacos , Resistencia a Antineoplásicos/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Mesilato de Imatinib/farmacología , Ratones , Mutación , Cromosoma Filadelfia , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Inhibidores de Proteínas Quinasas/farmacologíaRESUMEN
BACKGROUND: Systemic sclerosis (SSc) is a systemic autoimmune disease characterized by excessive fibrosis. FcγRIIB is a low-affinity receptor for the Fc fragment of IgG. FcγRIIB is expressed on the surface of various leukocyte subsets and signals negative feedback pathways to down-regulate B-cell antigen receptor signaling. OBJECTIVE: The aim of the present study was to investigate the role of FcγRIIB in the development of a murine bleomycin-induced scleroderma model. METHODS: The experimental fibrosis model was generated by the intradermal injection of bleomycin into wild-type (WT) and FcγRIIB-deficient (FcγRIIB-/-) mice. We histologically assessed skin and lung fibrosis as well as inflammatory cell infiltration. Cytokine and chemokine expression levels were measured with RT-PCR. RESULTS: The severity of fibrosis in the skin and lung was significantly worse in FcγRIIB-/- mice than in WT mice. In the skin of bleomycin-treated mice, the numbers of CD8+ T cells, F4/80+ macrophages, MPO+ neutrophils, NK1.1+NK cells, and B220+ B cells were significantly higher in FcγRIIB-/- mice than in WT mice. The expression of TNF-α and IL-1ß was significantly higher in FcγRIIB-/- mice than in WT mice as was the expression of ICAM-1, CXCL2, and CCL3 in the affected skin. An adoptive transfer of splenic leukocytes from FcγRIIB-/- mice into WT mice showed exacerbated skin and lung fibrosis compared to WT mice without an adoptive transfer. CONCLUSION: These results indicate that FcγRIIB plays an inhibitory role in skin and lung fibrosis. Moreover, modulating FcγRIIB signaling has potential as a therapeutic approach for SSc.
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Bleomicina , Esclerodermia Sistémica , Animales , Bleomicina/toxicidad , Linfocitos T CD8-positivos/metabolismo , Modelos Animales de Enfermedad , Fibrosis , Ratones , Esclerodermia Sistémica/patología , Piel/patologíaRESUMEN
Sphingosine-1-phosphate (S1P) regulates numerous important physiological functions, including immune response and vascular integrity, via its cognate receptors (S1PR1 to S1PR5); however, it remains unclear how S1P activates S1PRs upon binding. Here, we determined the crystal structure of the active human S1PR3 in complex with its natural agonist S1P at 3.2-Å resolution. S1P exhibits an unbent conformation in the long tunnel, which penetrates through the receptor obliquely. Compared with the inactive S1PR1 structure, four residues surrounding the alkyl tail of S1P (the "quartet core") exhibit orchestrating rotamer changes that accommodate the moiety, thereby inducing an active conformation. In addition, we reveal that the quartet core determines G protein selectivity of S1PR3. These results offer insight into the structural basis of activation and biased signaling in G protein-coupled receptors and will help the design of biased ligands for optimized therapeutics.
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Melanoma , Neoplasias Cutáneas , Protocolos de Quimioterapia Combinada Antineoplásica , Humanos , Imidazoles/uso terapéutico , Melanoma/tratamiento farmacológico , Mutación , Metástasis de la Neoplasia , Oximas/uso terapéutico , Proteínas Proto-Oncogénicas B-raf/genética , Piridonas/uso terapéutico , Pirimidinonas/uso terapéutico , Neoplasias Cutáneas/tratamiento farmacológicoAsunto(s)
Linfohistiocitosis Hemofagocítica/etiología , Micosis Fungoide/complicaciones , Neoplasias Cutáneas/complicaciones , Anciano , Médula Ósea/patología , Resultado Fatal , Femenino , Humanos , Ganglios Linfáticos/patología , Linfohistiocitosis Hemofagocítica/patología , Micosis Fungoide/patología , Neoplasias Cutáneas/patologíaRESUMEN
De novo formation of the double-membrane compartment autophagosome is seeded by small vesicles carrying membrane protein autophagy-related 9 (ATG9), the function of which remains unknown. Here we find that ATG9A scrambles phospholipids of membranes in vitro. Cryo-EM structures of human ATG9A reveal a trimer with a solvated central pore, which is connected laterally to the cytosol through the cavity within each protomer. Similarities to ABC exporters suggest that ATG9A could be a transporter that uses the central pore to function. Moreover, molecular dynamics simulation suggests that the central pore opens laterally to accommodate lipid headgroups, thereby enabling lipids to flip. Mutations in the pore reduce scrambling activity and yield markedly smaller autophagosomes, indicating that lipid scrambling by ATG9A is essential for membrane expansion. We propose ATG9A acts as a membrane-embedded funnel to facilitate lipid flipping and to redistribute lipids added to the outer leaflet of ATG9 vesicles, thereby enabling growth into autophagosomes.
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Autofagosomas/química , Proteínas Relacionadas con la Autofagia/química , Proteínas de la Membrana/química , Fosfolípidos/química , Proteolípidos/química , Proteínas de Transporte Vesicular/química , Animales , Autofagosomas/metabolismo , Proteínas Relacionadas con la Autofagia/genética , Proteínas Relacionadas con la Autofagia/metabolismo , Sitios de Unión , Transporte Biológico , Línea Celular , Microscopía por Crioelectrón , Fibroblastos/metabolismo , Fibroblastos/ultraestructura , Expresión Génica , Proteínas Fluorescentes Verdes/química , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Células HEK293 , Células HeLa , Humanos , Membrana Dobles de Lípidos/química , Membrana Dobles de Lípidos/metabolismo , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Ratones , Simulación de Dinámica Molecular , Fosfolípidos/metabolismo , Unión Proteica , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Multimerización de Proteína , Proteolípidos/metabolismo , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Proteínas de Transporte Vesicular/genética , Proteínas de Transporte Vesicular/metabolismo , Proteína Fluorescente RojaAsunto(s)
Adenosina Trifosfatasas/inmunología , Anticuerpos Antinucleares/sangre , Proteínas de Unión al ADN/inmunología , Dermatomiositis/sangre , Neoplasias Uterinas/cirugía , Creatina Quinasa/sangre , Dermatomiositis/complicaciones , Dermatomiositis/tratamiento farmacológico , Femenino , Fructosa-Bifosfato Aldolasa/sangre , Humanos , Persona de Mediana Edad , Periodo Posoperatorio , Neoplasias Uterinas/complicacionesRESUMEN
A 79-year-old man visited our hospital with a chief complaint of erythema around the urethral orifice of the glans, and a diagnosis of penile Bowen's disease was made by penile skin biopsy. Initially, local tumor resection with skin grafting from the femoral skin was performed. However, histopathological examination revealed a positive urethral margin of tumor. Since the degree of urethral lesion of penile Bowen's disease was unclear, the patient received glansectomy. Histopathological examination revealed a tumor lesion in the distal urethra, 7 mm from the external urethral orifice, and confirmed the complete resection of urethral Bowen's disease. The patient recovered well, with no definite recurrence of Bowen's disease up to 13 months after partial penectomy. Human papillomavirus 16 DNA was detected in the paraffin-embedded tumor sample, and in situ hybridization confirmed human papillomavirus 16 DNA locations in the tumor tissue. Immunohistochemistry revealed that p16-INK4a was widely expressed in the tumor tissue.
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Styrene maleic acid copolymers (SMA) form discoidal lipid nanoparticles (lipid nanodisks) that mimic plasma high-density lipoproteins. We have previously prepared and characterized lipid nanodisks composed of SMA and the neutral phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). In the present study, we tested whether the surface charges can alter the physicochemical and biological properties of lipid-SMA discoidal particles. Unlike the case of DMPC alone, addition of saline to the buffer was necessary to induce the formation of lipid-SMA complexes containing either 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) or 1,2-dimyristoyl-3-trimethylammonium-propane (DMTAP), with formation efficiency being dependent on the concentration of charged lipids. After purification, DMPG- or DMTAP-containing discoidal particles with an approximate size of 10 nm were obtained in a manner similar to DMPC alone. Although DMPG and DMTAP appeared to be similarly incorporated into the lipid nanodisks, the zeta potentials of both particles were comparable. That is, no significant differences were observed in the physicochemical properties between the lipid-SMA nanodisks. Compared to DMPC-SMA nanodisks, the uptake of DMPG or DMTAP-containing discoidal particles by RAW264 cells was increased for both particle types, whereas in MDA-MB-231 cells, only DMTAP-containing discoidal particle uptake was increased. In addition, fluorescence microscopy revealed that lipid-SMA nanodisks are localized adjacent to the plasma membrane of RAW264 cells but in MDA-MB-231 cells they accumulated in the center of the cell. Furthermore, these particles caused cytotoxicity in a cell-type dependent manner, with high toxicity in MDA-MB-231. These results raised the possibility that compositional alterations in lipid-SMA discoidal particles may modulate biological reactions in vivo.
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Lipoproteínas/química , Maleatos/química , Maleatos/metabolismo , Poliestirenos/química , Poliestirenos/metabolismo , Membrana Celular/química , Dimiristoilfosfatidilcolina/química , Gotas Lipídicas/química , Lipoproteínas/metabolismo , Nanopartículas/química , Fosfolípidos/química , Solubilidad , Estireno/químicaRESUMEN
AIM: Systemic sclerosis (SSc) is an autoimmune disease characterized by skin and lung fibrosis. Although SSc has a high mortality risk, an effective treatment for the disease has not been established yet. Mesenchymal stromal/stem cells (MSCs) are multipotential nonhematopoietic progenitor cells that have the ability to regulate immune responses. Adipose-derived stromal/stem cells (ASCs), one of the types of MSCs, have the advantage of accessibility and potent immunomodulatory effects when compared with other MSCs, such as bone marrow-derived MSCs. This study aimed to investigate the antifibrotic effect of ASCs in scleroderma mouse models, including bleomycin-induced scleroderma and sclerodermatous chronic graft-versus-host disease (Scl-cGVHD) models. METHOD: ASCs were intravenously administered to a bleomycin-induced scleroderma or Scl-cGVHD model on day 0. We compared the skin and lung fibrosis of scleroderma model mice between the ASC-treated group and control group. RESULTS: Administration of ASCs attenuated the skin and lung fibrosis of bleomycin-induced scleroderma and Scl-cGVHD model mice compared to that in the control mice. Immunohistochemical staining showed that ASCs suppressed the infiltration of CD4+ , CD8+ T cells and macrophages into the dermis of bleomycin model mice compared to that in control mice. In addition, ASCs attenuated the messenger RNA expression of collagen and fibrogenic cytokines, such as interleukin (IL)-6 and IL-13, in the skin of bleomycin model mice. ASCs also reduced the frequency of fibrogenic cytokine-producing CD4+ T cells and effector B cells in the spleen of bleomycin model mice. CONCLUSION: ASCs could prove to be a potential therapeutic agent for use in patients with SSc.