RESUMO
Nanoparticle-based drug delivery strategies have emerged as a crucial avenue for comprehensive sensorineural hearing loss treatment. Nevertheless, developing therapy vectors crossing both biological and cellular barriers has encountered significant challenges deriving from various external factors. Herein, the rational integration of gelatin nanoparticles (GNPs) with tetrahedral DNA nanostructures (TDNs) to engineer a distinct drug-delivery nanosystem (designed as TDN@GNP) efficiently enhances the biological permeability and cellular internalization, further resolving the dilemma of noise-induced hearing loss via loading epigallocatechin gallate (EGCG) with anti-lipid peroxidation property. Rationally engineering of TDN@GNP demonstrates dramatic alterations in the physicochemical key parameters of TDNs that are pivotal in cell-particle interactions and promote cellular uptake through multiple endocytic pathways. Furthermore, the EGCG-loaded nanosystem (TDN-EGCG@GNP) facilitates efficient inner ear drug delivery by superior permeability through the biological barrier (round window membrane), maintaining high drug concentration within the inner ear. The TDN-EGCG@GNP actively overcomes the cell membrane, exhibiting hearing protection from noise insults via reduced lipid peroxidation in outer hair cells and spiral ganglion neurons. This work exemplifies how integrating diverse vector functionalities can overcome biological and cellular barriers in the inner ear, offering promising applications for inner ear disorders.
Assuntos
Catequina , DNA , Gelatina , Perda Auditiva Provocada por Ruído , Nanoestruturas , Gelatina/química , DNA/química , DNA/metabolismo , Perda Auditiva Provocada por Ruído/metabolismo , Perda Auditiva Provocada por Ruído/tratamento farmacológico , Animais , Nanoestruturas/química , Catequina/análogos & derivados , Catequina/química , Catequina/farmacologia , Camundongos , Peroxidação de Lipídeos/efeitos dos fármacos , Nanopartículas/química , Sistemas de Liberação de MedicamentosRESUMO
Conductive metal-organic frameworks (cMOFs), which have high porosity and intrinsic electron conductivity, are regarded as ideal candidates for electromagnetic wave (EMW) absorption materials. Controlling the nanostructure of absorbers may be one of the effective strategies to improve the electromagnetic wave (EMW) absorption performance. Herein, a series of conductive Cu-HHTP MOFs (HHTP = 2,3,6,7,10,11-hexahydroxytriphenyl hydrates) with different nanostructures or crystal morphologies were successfully synthesized by using different structural inducers to regulate the changes in the morphology, thereby improving the EMW absorption performance. Specifically, when ammonia was used as an inducer, the obtained A-Cu-HHTP with a nanosheet structure exhibited excellent EMW absorption performance. The minimum reflection loss (RLmin) can reach -51.08 dB at 7.25 GHz with a thickness of 4.4 mm, and the maximum effective absorption bandwidth (EAB) can cover 5.73 GHz at 2.5 mm. The influence of the nanostructures of the cMOFs on the dielectric and EMW absorption performance was clarified. The nanosheet structure of A-Cu-HHTP increases its specific surface area, which expands multiple scattering and reflection paths of incident EMW; Meanwhile, the unique structure facilitates the formation of more heterogeneous interfaces, optimizing impedance matching. The significant improvement in EMW performance is mainly attributed to multiple reflections and scattering as well as impedance matching. This work not only provides a simple and effective strategy for improving electromagnetic wave absorption performance but also offers guidelines for preparing morphology functional cMOF materials.
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Otoferlin (OTOF) gene mutations represent the primary cause of hearing impairment and deafness in auditory neuropathy. The c.2485C>T (p. Q829X) mutation variant is responsible for approximately 3% of recessive prelingual deafness cases within the Spanish population. Previous studies have used two recombinant AAV vectors to overexpress OTOF, albeit with limited efficacy. In this study, we introduce an enhanced mini-dCas13X RNA base editor (emxABE) delivered via an AAV9 variant, achieving nearly 100% transfection efficiency in inner hair cells. This approach is aimed at treating OTOFQ829X, resulting in an approximately 80% adenosine-to-inosine conversion efficiency in humanized OtofQ829X/Q829X mice. Following a single scala media injection of emxABE targeting OTOFQ829X (emxABE-T) administered during the postnatal day 0-3 period in OtofQ829X/Q829X mice, we observed OTOF expression restoration in nearly 100% of inner hair cells. Moreover, auditory function was significantly improved, reaching similar levels as in wild-type mice. This enhancement persisted for at least 7 months. We also investigated P5-P7 and P30 OtofQ829X/Q829X mice, achieving auditory function restoration through round window injection of emxABE-T. These findings not only highlight an effective therapeutic strategy for potentially addressing OTOFQ829X-induced hearing loss but also underscore emxABE as a versatile toolkit for treating other monogenic diseases characterized by premature termination codons.
Assuntos
Surdez , Perda Auditiva Central , Perda Auditiva , Animais , Camundongos , Edição de Genes , Perda Auditiva/genética , Perda Auditiva/terapia , MutaçãoRESUMO
Cisplatin (DDP)-based chemotherapy is the main chemotherapeutic agent for ovarian cancer (OC) treatment. Circular RNA PIP5K1A (circ-PIP5K1A) was found to promote OC tumorigenesis. However, whether circ-PIP5K1A was involved in DDP resistance in OC remains unclear. Levels of circ-PIP5K1A, microRNA (miR)-942-5p, and nuclear factor I B (NFIB) were detected using quantitative real-time PCR and Western blot assays. In-vitro experiments were conducted by using cell counting kit-8, cell colony formation, 5-ethynyl-2'-deoxyuridine, flow cytometry, and transwell assays, respectively. In-vivo assay was performed using murine xenograft model. The binding interaction between miR-942-5p and circ-PIP5K1A or NFIB was confirmed using dual-luciferase reporter assay. Exosomes were obtained from culture media by the use of commercial kits and qualified by transmission electron microscopy and Western blot. Circ-PIP5K1A was highly expressed in DDP-resistant OC tissues and cells. Circ-PIP5K1A knockdown could constrain the proliferation, migration, and invasion, as well as increase apoptosis and sensitivity to DDP in DDP-resistant OC cells. Mechanistically, circ-PIP5K1A acted as a sponge for miR-942-5p to positively regulate NFIB expression. Moreover, rescue experiments demonstrated that the anticancer and DDP sensitization effects caused by circ-PIP5K1A silencing in DDP-resistant OC cells were achieved through the miR-942-5p/NFIB axis. Importantly, circ-PIP5K1A silencing enhanced DDP efficacy and impeded tumor growth in OC in vivo . Additionally, we also found that circ-PIP5K1A was packaged into exosomes and could be internalized by surrounding cells. Circ-PIP5K1A knockdown reduced the resistance to DDP in OC via regulating miR-942-5p/NFIB axis. Besides that, circ-PIP5K1A was packaged into exosomes and exosomal circ-SKA3 could mediate intercellular communication between OC cells. These findings provided a promising therapeutic target for OC.
Assuntos
MicroRNAs , Neoplasias Ovarianas , Animais , Feminino , Humanos , Camundongos , Proteínas de Ciclo Celular , Proliferação de Células , Cisplatino , Resistencia a Medicamentos Antineoplásicos , Proteínas Associadas aos Microtúbulos , Fatores de Transcrição NFI , RNA CircularRESUMO
BACKGROUND: Rituximab (RTX) is a standard therapy for antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). However, the most frequently used dose may lead to severe adverse effects (SAEs). We explored the efficacy and safety of low-dose RTX in Chinese patients with AAV. METHODS: A total of 22 Chinese patients diagnosed with AAV with renal involvement, including 8 treated with low-dose RTX (400 mg of RTX total over 4 weeks) and 14 treated with cyclophosphamide (CYC), were evaluated. The baseline clinical and pathological data and laboratory parameters during follow-up at months 1, 3, 6, and 12 were collected retrospectively. RESULTS: The baseline data showed no significant differences between the two groups. The median peripheral CD19+ cell counts in the RTX group decreased from 315.0/µL to 1.5/µL at 2 weeks, and to 2.5/µL at 1 month after the first dose. The median SCr level decreased from 267.8 µmol/L before treatment to 151.45 µmol/L at 1 month, 132.75 µmol/L at 3 months, 123.2 µmol/L at 6 months, and 151.9 µmol/L at 12 months in RTX-treated patients. The improvements in renal function, proteinuria, and ANCA titre were not significantly different between the two groups. The SAE rate was significantly lower in the RTX group (one SAE of pneumonia) compared with the CYC group. CONCLUSIONS: This is the first report that low-dose RTX could be effective for the treatment of Chinese patients with AAV with renal involvement.
Assuntos
Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos , Anticorpos Anticitoplasma de Neutrófilos , Rituximab , Humanos , Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos/diagnóstico , Ciclofosfamida/efeitos adversos , Ciclofosfamida/uso terapêutico , População do Leste Asiático , Quimioterapia de Indução , Indução de Remissão , Estudos Retrospectivos , Rituximab/efeitos adversos , Rituximab/uso terapêutico , Resultado do TratamentoRESUMO
PURPOSE: This study aimed to quantitatively assess the incidence of hearing loss in relation to age in individuals with biallelic p.V37I variant in GJB2. METHODS: Population screening of the biallelic p.V37I variant was performed in 30,122 individuals aged between 0 and 97 years in Shanghai. Hearing thresholds of the biallelic p.V37I individuals and the controls were determined by click auditory brainstem response or pure tone audiometry. RESULTS: Biallelic p.V37I was detected in 0.528% (159/30,122) of the subjects. Of the biallelic p.V37I newborns, 43.91% (18/41) passed their distortion-product otoacoustic emissions-based newborn hearing screening or had hearing thresholds lower than 20 decible above normal hearing level. The older newborns had elevated hearing thresholds, with increasing incidence of 9.52%, 23.08%, 59.38%, and 80.00% for moderate or higher grade of hearing loss in age groups of 7 to 15 years, 20 to 40 years, 40 to 60 years, and 60 to 85 years, respectively. Their hearing deteriorated at a rate of 0.40 dB hearing level per year on average; males were more susceptible, and deterioration occurred preferentially at higher sound frequencies. CONCLUSION: The biallelic p.V37I variant is associated with steadily progressive hearing loss with increasing incidence over the course of life. Most of the biallelic p.V37I individuals may develop significant hearing loss in adulthood and, can benefit from early diagnosis and intervention through wide-spread genetic screening.
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Surdez , Perda Auditiva , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , China/epidemiologia , Conexina 26 , Conexinas/genética , Surdez/genética , Perda Auditiva/diagnóstico , Perda Auditiva/epidemiologia , Perda Auditiva/genética , Humanos , Incidência , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
BACKGROUND: Excessive accumulation of reactive oxygen species (ROS) has been documented as the crucial cellular mechanism of cisplatin-induced ototoxicity. However, numerous antioxidants have failed in clinical studies partly due to inefficient drug delivery to the cochlea. A drug delivery system is an attractive strategy to overcome this drawback. METHODS AND RESULTS: In the present study, we proposed the combination of antioxidant astaxanthin (ATX) and ROS-responsive/consuming nanoparticles (PPS-NP) to combat cisplatin-induced ototoxicity. ATX-PPS-NP were constructed by the self-assembly of an amphiphilic hyperbranched polyphosphoester containing thioketal units, which scavenged ROS and disintegrate to release the encapsulated ATX. The ROS-sensitivity was confirmed by 1H nuclear magnetic resonance spectroscopy, transmission electron microscopy and an H2O2 ON/OFF stimulated model. Enhanced release profiles stimulated by H2O2 were verified in artificial perilymph, the HEI-OC1 cell line and guinea pigs. In addition, ATX-PPS-NP efficiently inhibited cisplatin-induced HEI-OC1 cell cytotoxicity and apoptosis compared with ATX or PPS-NP alone, suggesting an enhanced effect of the combination of the natural active compound ATX and ROS-consuming PPS-NP. Moreover, ATX-PPS-NP attenuated outer hair cell losses in cultured organ of Corti. In guinea pigs, NiRe-PPS-NP verified a quick penetration across the round window membrane and ATX-PPS-NP showed protective effect on spiral ganglion neurons, which further attenuated cisplatin-induced moderate hearing loss. Further studies revealed that the protective mechanisms involved decreasing excessive ROS generation, reducing inflammatory chemokine (interleukin-6) release, increasing antioxidant glutathione expression and inhibiting the mitochondrial apoptotic pathway. CONCLUSIONS: Thus, this ROS-responsive nanoparticle encapsulating ATX has favorable potential in the prevention of cisplatin-induced hearing loss.
Assuntos
Antineoplásicos , Perda Auditiva , Nanopartículas , Ototoxicidade , Animais , Antineoplásicos/uso terapêutico , Antioxidantes/uso terapêutico , Apoptose , Cisplatino/farmacologia , Cobaias , Perda Auditiva/induzido quimicamente , Perda Auditiva/tratamento farmacológico , Perda Auditiva/prevenção & controle , Peróxido de Hidrogênio , Nanopartículas/química , Espécies Reativas de Oxigênio/metabolismo , XantofilasRESUMO
Recent studies have observed that lncRNAs (long non-coding RNAs) are involved in the progression of various tumours including tongue squamous cell carcinoma (TSCC). Recently, a new lnRNA, GACAT1, has been firstly identified in gastric cancer. However, its potential role in TSCC remains unknown. In this reference, we observed that GACAT1 was overexpressed in TSCC samples and cell lines. Of 25 TSCC specimens, GACAT1 expression was overexpressed in 18 patients (18/25, 72%) compared to non-tumour specimens. Ectopic expression of GACAT1 induced cell growth and migration and promoted epithelial to mesenchymal transition in TSCC. In addition, ectopic expression of GACAT1 decreased miR-149 expression in SCC1 cell. We observed that miR-149 expression was down-regulated in TSCC cell lines. Moreover, we observed that GACAT1 expression was negatively correlated with miR-149 expression. GACAT1 overexpression induced TSCC cell growth and migration via regulating miR-149 expression. These data provided that GACAT1 played an oncogenic role in the progression of TSCC partly through modulating miR-149 expression.
Assuntos
Carcinoma de Células Escamosas/metabolismo , MicroRNAs/metabolismo , RNA Longo não Codificante/fisiologia , Neoplasias da Língua/metabolismo , Estudos de Casos e Controles , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , HumanosRESUMO
Homochiral metal-organic frameworks (HMOFs) have garnered considerable attention due to their extrachiral properties and broad application for chiral recognition. However, assembling a pair of high-quality chiral MOFs for sensing enantiomers precisely is a formidable challenge because of the complicated chiral environment and uncontrollable coordinated conditions. Herein, one pair of homochiral UiO-66 analogues, S-1 (l-AP@UiO-66-(COOH)2) and R-1 (d-AP@UiO-66-(COOH)2), are reported for chiral recognition. They were fabricated via a condensation reaction between the carboxyl groups of UiO-66-(COOH)2 and amino groups of l/d-amino propanol (l/d-AP). These novel fluorescent probes exhibited highly enantioselective fluorescence enhancement towards l/d-phenylalaninol (l/d-PA). For example, when S-1 and R-1 were treated with l-PA or d-PA, they displayed different fluorescence responses: the enantiomeric fluorescence enhancement ratio (ef) was 2.51 and 0.41 for S-1 and R-1, respectively. Hence, a visible difference in fluorescence enhancement for l-PA and d-PA and excellent enantioselective behavior between S-1 and l-PA (or R-1 and d-PA) was displayed. Measurements of fluorescence lifetime, powder X-ray diffraction, molecular-dynamic simulations and Benesi-Hildebrand plots were employed to determine the observed high enantioselectivity for l/d-PA. In brief, we found that two post-modified HMOFs, S-1 and R-1, were outstanding enantioselective sensors for detecting l-PA and d-PA. They had a prominent difference in ef and remarkable enantioselectivity factor α and ΔΔG based on steric hindrance and stereochemical difference.
Assuntos
Corantes Fluorescentes , Fenilalanina , Fluorescência , Fenilalanina/análogos & derivados , EstereoisomerismoRESUMO
It is widely accepted that even a single acute noise exposure at moderate intensity that induces temporary threshold shift (TTS) can result in permanent loss of ribbon synapses between inner hair cells and afferents. However, effects of repeated or chronic noise exposures on the cochlear synapses especially medial olivocochlear (MOC) efferent synapses remain elusive. Based on a weeklong repeated exposure model of bandwidth noise over 2-20 kHz for 2 hours at seven intensities (88 to 106 dB SPL with 3 dB increment per gradient) on C57BL/6J mice, we attempted to explore the dose-response mechanism of prolonged noise-induced audiological dysfunction and cochlear synaptic degeneration. In our results, mice repeatedly exposed to relatively low-intensity noise (88, 91, and 94 dB SPL) showed few changes on auditory brainstem response (ABR), ribbon synapses, or MOC efferent synapses. Notably, repeated moderate-intensity noise exposures (97 and 100 dB SPL) not only caused hearing threshold shifts and the inner hair cell ribbon synaptopathy but also impaired MOC efferent synapses, which might contribute to complex patterns of damages on cochlear function and morphology. However, repeated high-intensity (103 and 106 dB SPL) noise exposures induced PTSs mainly accompanied by damages on cochlear amplifier function of outer hair cells and the inner hair cell ribbon synaptopathy, rather than the MOC efferent synaptic degeneration. Moreover, we observed a frequency-dependent vulnerability of the repeated acoustic trauma-induced cochlear synaptic degeneration. This study provides a sight into the hypothesis that noise-induced cochlear synaptic degeneration involves both afferent (ribbon synapses) and efferent (MOC terminals) pathology. The pattern of dose-dependent pathological changes induced by repeated noise exposure at various intensities provides a possible explanation for the complicated cochlear synaptic degeneration in humans. The underlying mechanisms remain to be studied in the future.
Assuntos
Perda Auditiva Provocada por Ruído/etiologia , Vias Aferentes/fisiopatologia , Animais , Vias Auditivas/fisiologia , Limiar Auditivo , Cóclea , Vias Eferentes/fisiopatologia , Células Ciliadas Auditivas Internas/fisiologia , Perda Auditiva Provocada por Ruído/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Doenças Neurodegenerativas/etiologia , Doenças Neurodegenerativas/fisiopatologia , Núcleo Olivar/fisiologia , Recidiva , SinapsesRESUMO
Due to the disruption by other nonanalyte factors, single-emission probes have been limited in complicated detecting systems. In this work, a pH-modulated luminescence chameleon system based on lanthanide-based MOF (Eu3+@Mn-MOF), with stable structure and miraculous dual-emitting fluorescent properties, was synthesized by a postsynthetic modification (PSM) strategy of a simple hydrothermal and agitation method. Amazingly, not only can the Eu3+@Mn-MOF emit a broad emission at 500 nm attributed to the ligand-based fluorescence emission but it can also exhibit the characteristic emission of Eu3+ ions responding to the antenna effect. Moreover, the Eu3+@Mn-MOF displays an interesting luminescence color transition between acidic and basic solutions. Inspired by this phenomenon, a pH-modulated luminescence chameleon system was first constructed and employed to detect histidine, a kind of basic amino acid for a variety of biological matters, causing a unique fluorescence signal of the ratio-dependent color to change from yellow to light pink which differs from the color change of other water-soluble amino acids. Therefore, Eu3+@Mn-MOF can be as a practical pH-modulated luminescence chameleon system chemsensor for sensing histidine with low detection limit, high sensitivity, and rapid sensing time. In conclusion, the postsynthetic modified Eu3+@Mn-MOF has outstanding applications in the fields of chemical detection and human health.
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Európio/química , Histidina/análise , Luminescência , Substâncias Luminescentes/química , Manganês/química , Estruturas Metalorgânicas/química , Concentração de Íons de Hidrogênio , Substâncias Luminescentes/síntese química , Medições Luminescentes , Estruturas Metalorgânicas/síntese químicaRESUMO
An amendment to this paper has been published and can be accessed via the original article.
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BACKGROUND: Ototoxicity is one of the major side effects of platinum-based chemotherapy, especially cisplatin therapy. To date, no FDA approved agents to alleviate or prevent this ototoxicity are available. However, ototoxicity is generally believed to be produced by excessive generation of reactive oxygen species (ROS) in the inner ear, thus leading to the development of various antioxidants, which act as otoprotective agents. Astaxanthin (ATX) is an interesting candidate in the development of new therapies for preventing and treating oxidative stress-related pathologies, owing to its unique antioxidant capacity. METHODS AND RESULTS: In this study, we aimed to evaluate the potential antioxidant properties of ATX in the inner ear by using the HEI-OC1 cell line, zebrafish, and guinea pigs. Because ATX has poor solubility and cannot pass through round window membranes (RWM), we established lipid-polymer hybrid nanoparticles (LPN) for loading ATX. The LPN enabled ATX to penetrate RWM and maintain concentrations in the perilymph in the inner ear for 24 h after a single injection. ATX-LPN were found to have favorable biocompatibility and to strongly affect cisplatin-induced generation of ROS, on the basis of DCFHDA staining in HEI-OC1 cells. JC-1 and MitoTracker Green staining suggested that ATX-LPN successfully reversed the decrease in mitochondrial membrane potential induced by cisplatin in vitro and rescued cells from early stages of apoptosis, as demonstrated by FACS stained with Annexin V-FITC/PI. Moreover, ATX-LPN successfully attenuated OHC losses in cultured organ of Corti and animal models (zebrafish and guinea pigs) in vivo. In investigating the protective mechanism of ATX-LPN, we found that ATX-LPN decreased the expression of pro-apoptotic proteins (caspase 3/9 and cytochrome-c) and increased expression of the anti-apoptotic protein Bcl-2. In addition, the activation of JNK induced by CDDP was up-regulated and then decreased after the administration of ATX-LPN, while P38 stayed unchanged. CONCLUSIONS: To best of our knowledge, this is first study concluded that ATX-LPN as a new therapeutic agent for the prevention of cisplatin-induced ototoxicity.
Assuntos
Nanopartículas/química , Polímeros/farmacologia , Xantofilas/farmacologia , Animais , Antioxidantes , Apoptose/efeitos dos fármacos , Caspase 3 , Linhagem Celular , Cisplatino/farmacologia , Orelha Interna/metabolismo , Cobaias , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Modelos Animais , Polímeros/química , Proteínas Proto-Oncogênicas c-bcl-2 , Espécies Reativas de Oxigênio/metabolismo , Xantofilas/química , Peixe-ZebraRESUMO
Genetic hearing impairment is highly heterogeneous. In this study, targeted next-generation sequencing (NGS) in two Chinese Han families identified a novel p.G141R homozygous mutation in ILDR1 as the genetic cause of the deafness. Consistent with the recessive inheritance, cosegregation of the p.G141R variant with the hearing loss was confirmed in members of both families by PCR amplification and Sanger sequencing. SNP genotyping analysis suggested that those two families were not closely related. Our study showed that targeted NGS is an effective tool for diagnosis of genetic deafness and that p.G141R in ILDR1 may be a relatively frequent mutation for DFNB42 in Chinese Hans.
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Surdez/genética , Predisposição Genética para Doença , Perda Auditiva Neurossensorial/genética , Receptores de Superfície Celular/genética , Povo Asiático/genética , China , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Mutação , Linhagem , Polimorfismo de Nucleotídeo ÚnicoRESUMO
PURPOSE: Prostate cancer (PCa) is the most frequently malignant neoplasm in men. MicroRNAs (miRs) have been identified to play important biological roles in a variety of tumors. Several studies showed that miR-191 was involved in the development of different cancers, but its role in prostate cancer remains unclear. METHODS: Human PCa cell lines DU145, PC-3 and LNCAP, and benign prostate hyperplasia (BPH) and human prostate epithelial cell line RWPE-1 were used. The expression level of miR-191 in 48 paired prostate tumor and adjacent normal tissues was assessed along with the clinical patient features. Synthetic miR-191 mimics and inhibitors were used to overexpress or inhibit the miR-191 level. CCK8 and colony formation assay were used to evaluate the cell growth. The ability of cell invasion was studied by transwell assay. Dual-luciferase experiment was used to identify the target gene and western blot was performed to evaluate the protein level. RESULTS: miR-191 was overexpressed in PCa tissue samples compared to the normal group as well in PCa-derived cell lines. Upregulation miR-191 in PC-3 cells significantly promoted while downregulation miR-191 in DU145 cells retarded the cell proliferation and invasion. Furthermore, TIMP3 were proved to be a direct target gene of miR-191 and knockdown of TIMP3 reversed the function of miR-191 downregulation. CONCLUSION: This study demonstrated that miR-191 promoted the cell growth and invasion ability in prostate cancer through downregulating TIMP3 and might be a potential target for the biotherapy for PCa.
Assuntos
MicroRNAs/genética , Hiperplasia Prostática/genética , Neoplasias da Próstata/genética , Inibidor Tecidual de Metaloproteinase-3/genética , Idoso , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Próstata , Hiperplasia Prostática/patologia , Neoplasias da Próstata/patologiaRESUMO
PURPOSE: To explore the genetic etiology of deafness in a dominant family with late-onset, progressive, nonsyndromic hearing loss. METHODS: Genome-wide linkage analysis was performed for 21 family members. Candidate pathogenic variants were identified by whole-exome sequencing of selected family members and confirmed by Sanger sequencing of all family members. Cochlear expression of Dmxl2 was investigated by reverse-transcription polymerase chain reaction (RT-PCR) and immunostaining of the organ of Corti from mice. RESULTS: The causative gene was mapped to a 9.68-Mb candidate region on chromosome 15q21.2 (maximum logarithm of the odds score = 4.03) that contained no previously described deafness genes. Whole-exome sequencing identified heterozygous c.7250G>A (p.Arg2417His) in DMXL2 as the only candidate pathogenic variant segregating the hearing loss. In mouse cochlea, expression of DMXL2 was restricted to the hair cells and the spiral ganglion neurons. CONCLUSION: Our data indicated that the p.Arg2417His variant in DMXL2 is associated with dominant, nonsyndromic hearing loss and suggested an important role of DMXL2 in inner ear function.Genet Med advance online publication 22 September 2016.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Surdez/genética , Mutação de Sentido Incorreto , Proteínas do Tecido Nervoso/genética , Órgão Espiral/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Idade de Início , Animais , China/etnologia , Surdez/metabolismo , Feminino , Estudos de Associação Genética , Ligação Genética , Predisposição Genética para Doença , Humanos , Masculino , Camundongos , Proteínas do Tecido Nervoso/metabolismo , Linhagem , Mapeamento Físico do Cromossomo , Sequenciamento do ExomaRESUMO
We investigated the role of serum response factor (SRF) in epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells (TECs) in diabetic nephropathy (DN). The expression of SRF, epithelial markers (E-cadherin and ZO-1), and mesenchymal markers (fibronectin, collagen-1, α-SMA, FSP-1) was examined in human proximal renal tubular epithelial cells (HK-2 cells) or renal medulla tissues following high glucose. SRF was upregulated by SRF plasmids and downregulated by CCG-1423 (a small molecule inhibitor of SRF) to investigate how SRF influenced EMT in TECs of DN. Streptozotocin was used to generate DM in rats. In HK-2 cells after high-glucose treatment and renal medulla tissues of diabetic rats, SRF, fibronectin, collagen-1, α-SMA, and FSP-1 increased, while E-cadherin and ZO-1 declined. SRF overexpression in HK-2 cells induced expression of Snail, an important transcription factor mediating EMT. Blockade of SRF by CCG-1423 reduced Snail induction and protected TECs from EMT both in vitro and in vivo. Together, increased SRF activity promotes EMT in TECs and dysfunction in DN. Targeting SRF by small molecule inhibitor may be an attractive therapeutic strategy for DN.
Assuntos
Nefropatias Diabéticas/metabolismo , Transição Epitelial-Mesenquimal/fisiologia , Túbulos Renais/metabolismo , Fatores de Transcrição/metabolismo , Animais , Linhagem Celular , Diabetes Mellitus Experimental/metabolismo , Regulação para Baixo/fisiologia , Células Epiteliais/metabolismo , Glucose/metabolismo , Masculino , Ratos , Ratos Wistar , Transdução de Sinais/fisiologia , Regulação para Cima/fisiologiaRESUMO
We investigated the expression and function of serum response factor (SRF) in endothelial-mesenchymal transition (EndMT) in glomerular endothelial cells (GEnCs) of diabetic nephropathy (DN). The expression of SRF, endothelial markers (VE-cadherin, CD31), and mesenchymal markers (α-SMA, FSP-1, fibronectin) was examined in GEnCs following high glucose or in renal cortex tissues of DN rats. SRF was upregulated by SRF plasmids and downregulated by CCG-1423 (a small molecule inhibitor of SRF) to investigate how SRF influenced EndMT in GEnCs of DN. Streptozocin (STZ) was used to generate diabetes mellitus DM in rats. In GEnCs after high glucose treatment and in renal cortex tissues of diabetic rats, SRF, α-SMA, FSP-1, and fibronectin increased, while VE-cadherin and CD31 declined. SRF overexpression in GEnCs induced expression of Snail, an important transcription factor mediating EndMT. Blockade of SRF reduced Snail induction, protected GEnCs from EndMT, and ameliorated proteinuria. Together, increased SRF activity provokes EndMT and barrier dysfunction of GEnCs in DN. Targeting SRF by small molecule inhibitor may be an attractive therapeutic strategy for DN.
Assuntos
Nefropatias Diabéticas/metabolismo , Células Endoteliais/metabolismo , Transição Epitelial-Mesenquimal/fisiologia , Proteinúria/metabolismo , Fatores de Transcrição/metabolismo , Animais , Biomarcadores/metabolismo , Linhagem Celular , Diabetes Mellitus Experimental/metabolismo , Regulação para Baixo/fisiologia , Glucose/metabolismo , Masculino , Ratos , Ratos Wistar , Transdução de Sinais/fisiologia , Regulação para Cima/fisiologiaRESUMO
Xiyanping is used to treat infectious diseases with antibiotics in clinic. The aim of this study is to investigate the mechanism of Xiyanping through studying the effect of the combination of Xiyanping with Cefazolin on the chemotaxis and phagocytic function of peripheral blood neutrophils in mice. Ten healthy mice were in control group. Forty healthy mice in experimental group were infected with staphylococcus aureus, and were randomly divided further into four groups, i. e. model group, Xiyanping group, Cefazolin group and combination group (Xiyanping with Cefazolin). Mice in the control group and model group were given normal saline (NS) through abdomen while those in other groups were given Xiyanping, Cefazolin, and Xiyanping with Cefazolin, respectively. The chemotaxis of peripheral blood neutrophils was detected with the transwell method, and the phagocytic function of peripheral blood neutrophils was analyzed with flow cytometry (FCM). In the present study, there was no significance on the chemotactic index of peripheral blood neutrophils in all the groups (P > 0.05). The actual phagocytotic rate and index of peripheral blood neutrophils in the blank group, Xiyanping group, and the combination group were significantly higher than those of the model group and Cefazolin group (P < 0.05). However, those were not significant in the blank group, Xiyanping group, and the combination group (P > 0.05) or between the model group and Cefazolin group (P> 0.05). Our results suggested the combination of Xiyanping and Cefazolin could enhance the therapeutic effect by improving the phagocytic function of peripheral blood neutrophils.
Assuntos
Antibacterianos/farmacologia , Cefazolina/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Neutrófilos/efeitos dos fármacos , Animais , Quimiotaxia , Modelos Animais de Doenças , Camundongos , Neutrófilos/citologia , Fagocitose , Infecções Estafilocócicas/imunologia , Staphylococcus aureusRESUMO
Clarifying the relationship between soil microorganisms and the plant-soil system is crucial for encouraging the sustainable development of ecosystems, as soil microorganisms serve a variety of functional roles in the plant-soil system. In this work, the influence mechanisms of significant soil microbial groups on the plant-soil system and their applications in environmental remediation over the previous 30 years were reviewed using a systematic literature review (SLR) methodology. The findings demonstrated that: (1) There has been a general upward trend in the number of publications on significant microorganisms, including bacteria, fungi, and archaea. (2) Bacteria and fungi influence soil development and plant growth through organic matter decomposition, nitrogen, phosphorus, and potassium element dissolution, symbiotic relationships, plant growth hormone production, pathogen inhibition, and plant resistance induction. Archaea aid in the growth of plants by breaking down low-molecular-weight organic matter, participating in element cycles, producing plant growth hormones, and suppressing infections. (3) Microorganism principles are utilized in soil remediation, biofertilizer production, denitrification, and phosphorus removal, effectively reducing environmental pollution, preventing soil pathogen invasion, protecting vegetation health, and promoting plant growth. The three important microbial groups collectively regulate the plant-soil ecosystem and help maintain its relative stability. This work systematically summarizes the principles of important microbial groups influence plant-soil systems, providing a theoretical reference for how to control soil microbes in order to restore damaged ecosystems and enhance ecosystem resilience in the future.