RESUMO
Oxylipins are important biological regulators that have received extensive research attention. Due to the extremely low concentrations, large concentration variations, and high structural similarity of many oxylipins, the quantitative analysis of oxylipins in biological samples is always a great challenge. Here, we developed a liquid chromatography-tandem mass spectrometry-based method with high sensitivity, wide linearity, and acceptable resolution for quantitative profiling of oxylipins in multiple biological samples. A total of 104 oxylipins, some with a high risk of detection crosstalk, were well separated on a 150 mm column over 20 min. The method showed high sensitivity with lower limits of quantitation for 87 oxylipins, reaching 0.05-0.5 pg. Unexpectedly, we found that the linear range for 16, 18, and 17 oxylipins reached 10,000, 20,000, and 40,000 folds, respectively. Due to the high sensitivity, while reducing sample consumption to below half the volume of previous methods, 74, 78, and 59 low-abundance oxylipins, among which some were difficult to detect like lipoxins and resolvins, were well quantified in the tested mouse plasma, mouse liver, and human plasma samples, respectively. Additionally, we determined that analytes with multifarious concentrations of over a 1,000-fold difference could be well quantified simultaneously due to the wide linearity. In conclusion, most likely due to the instrumental advancement, this method effectively improves the quantitative sensitivity and linear range over existing methods, which will facilitate and advance the study of the physiological and pathophysiological functions of oxylipins.
Assuntos
Oxilipinas , Espectrometria de Massas em Tandem , Humanos , Animais , Camundongos , Cromatografia Líquida , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida de Alta Pressão/métodos , Ácidos GraxosRESUMO
To minimize the gastric and esophageal injury effect, a system to deliver doxycycline hyclate (DOXY) to the duodenum area is needed. DOXY-containing modified-release oral pellets (DMOP) coated with hydroxypropyl methylcellulose phthalate HP-55 (HPMCP HP-55) and hydroxypropyl methylcellulose E15 (HPMC E15) appear to be a reasonable choice. This coating layer dissolves at pH 5.5, which is the pH of the duodenum, but not at a gastric pH (1.2). The formulation and preparation of DMOP were optimized, and a scale-up test was performed. The results showed that the production reproducibility was acceptable, and the quality of DMOP well met the standards of Chinese Pharmacopeia (Ch.P, 2015 edition). Notably, the accumulated DOXY release was lower than 50% at pH 1.2 (20 min) and higher than 85% at pH 5.5, which met the USP40-NF35 standard for DOXY modified-release formulations. Moreover, the storage stability of DMOP with different packages was investigated by stress testing, accelerated and long-term testings. The stability of DMOP was maintained up to 12 months, in terms of DOXY content and in vitro release behavior. The results seem to suggest that DMOP could be a promising duodenum delivery system.
Assuntos
Doxiciclina/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Administração Oral , Implantes de Medicamento/administração & dosagem , Implantes de Medicamento/química , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Concentração de Íons de Hidrogênio , Derivados da Hipromelose/química , Metilcelulose/análogos & derivados , Metilcelulose/química , Tamanho da Partícula , Comprimidos com Revestimento Entérico/administração & dosagem , Comprimidos com Revestimento Entérico/químicaRESUMO
1. Our objective is to investigate the alterations of hepatic drug transporters and metabolizing enzymes in hypercholesterolemia. Male Sprague-Dawley rats were fed high-cholesterol chows for 8 weeks to induce hypercholesterolemia. Protein levels of hepatic drug transporters and metabolizing enzymes were analyzed by iTRAQ labeling coupled with LC TRIPLE-TOF. 2. Total 239 differentially expressed proteins were identified using proteomic analysis. Among those, protein levels of hepatic drug transporters (MRP2, ABCD3, OAT2, SLC25A12, SCL38A3, SLC2A2 and SLC25A5) and metabolizing enzymes (CYP2B3, CYP2C7, CYP2C11, CYP2C13, CYP4A2 and UGT2B) were markedly reduced, but the levels of CYP2C6 and CYP2E1 were increased in hypercholesterolemia group compared to control. Decreased expressions of drug transporters MRP2 and OAT2 were further confirmed by real time quantitative PCR (RT-qPCR) and western blot. 3. Ingenuity pathway analysis revealed that these differentially expressed proteins were regulated by various signaling pathways including nuclear receptors and inflammatory cytokines. One of the nuclear receptor candidates, liver X receptor alpha (LXRα), was further validated by RT-qPCR and western blot. Additionally, LXRα agonist T0901317 rescued the reduced expressions of MRP2 and OAT2 in HepG2 cells in hypercholesterolemic serum treatment. 4. Our present results indicated that hypercholesterolemia affected the expressions of various drug transporters and metabolizing enzymes in liver via nuclear receptors pathway. Especially, decreased function of LXRα contributes to the reduced expressions of MRP2 and OAT2.
Assuntos
Hipercolesterolemia/metabolismo , Fígado/metabolismo , Proteoma/metabolismo , Transportadores de Cassetes de Ligação de ATP/metabolismo , Translocador 2 do Nucleotídeo Adenina/metabolismo , Animais , Hidrocarboneto de Aril Hidroxilases/metabolismo , Transporte Biológico , Sistema Enzimático do Citocromo P-450/metabolismo , Família 2 do Citocromo P450/metabolismo , Transportador de Glucose Tipo 2/metabolismo , Glucuronosiltransferase/metabolismo , Masculino , Proteínas dos Microfilamentos/metabolismo , Antígenos de Histocompatibilidade Menor/metabolismo , Proteínas de Transporte da Membrana Mitocondrial/metabolismo , Transportadores de Ânions Orgânicos Sódio-Independentes/metabolismo , Ratos , Ratos Sprague-Dawley , Esteroide 16-alfa-Hidroxilase/metabolismoRESUMO
The nontoxic heat-labile toxin (LT) B subunit (LTB) was used as mucosal adjuvant experimentally. However, the mechanism of LTB adjuvant was still unclear. The LTB and enterovirus 71 (EV71) VP1 subunit (EVP1) were constructed in pET32 and expressed in E. coli BL21, respectively. The immunogenicity of purified EVP1 and the adjuvanticity of LTB were evaluated via intranasal immunization EVP1 plus LTB in Balb/c mice. In order to elucidate the proteome change triggered by the adjuvant of LTB, the proteomic profiles of LTB, EVP1, and LTB plus EVP1 were quantitatively analyzed by iTRAQ-LC-MS/MS (isobaric tags for relative and absolute quantitation; liquid chromatography-tandem mass spectrometry) in murine macrophage RAW264.7. The proteomic data were analyzed by bioinformatics and validated by western blot analysis. The predicted protein interactions were confirmed using LTB pull-down and the LTB processing pathway was validated by confocal microscopy. The results showed that LTB significantly boosted EVP1 specific systematic and mucosal antibodies. A total of 3666 differential proteins were identified in the three groups. Pathway enrichment of proteomic data predicted that LTB upregulated the specific and dominant MAPK (mitogen-activated protein kinase) signaling pathway and the protein processing in endoplasmic reticulum (PPER) pathway, whereas LTB or EVP1 did not significantly upregulate these two signaling pathways. Confocal microscopy and LTB pull-down assays confirmed that the LTB adjuvant was endocytosed and processed through endocytosis (ENS)-lysosomal-endoplasmic reticulum (ER) system.
Assuntos
Toxinas Bacterianas/imunologia , Toxinas Bacterianas/metabolismo , Enterotoxinas/imunologia , Enterotoxinas/metabolismo , Enterovirus Humano A/metabolismo , Proteínas de Escherichia coli/imunologia , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Proteômica/métodos , Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/genética , Adjuvantes Imunológicos/metabolismo , Animais , Anticorpos Antivirais/imunologia , Toxinas Bacterianas/química , Toxinas Bacterianas/genética , Enterotoxinas/química , Enterotoxinas/genética , Enterovirus Humano A/genética , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Camundongos , Microscopia ConfocalRESUMO
O-GlcNAcylation is a dynamic and reversible posttranslational modification of nuclear and cytoplasmic proteins. In recent years, the roles of O-GlcNAcylation in several human malignant tumors have been investigated, and O-GlcNAcylation was found to be linked to cellular features relevant to metastasis. In this study, we modeled four diverse ovarian cancer cells and investigated the effects of O-GlcNAcylation on ovarian cancer cell migration. We found that total O-GlcNAcylation level was elevated in HO-8910PM cells compared to OVCAR3 cells. Additionally, through altering the total O-GlcNAcylation level by OGT silencing or OGA inhibition, we found that the migration of OVCAR3 cells was dramatically enhanced by PUGNAc and Thiamet G treatment, and the migration ability of HO-8910PM cells was significantly inhibited by OGT silencing. Furthermore, we also found that the expression of E-cadherin, an O-GlcNAcylated protein in ovarian cancer cells, was reduced by OGA inhibition in OVCAR3 cells and elevated by OGT silencing in HO-8910PM cells. These results indicate that O-GlcNAcylation could enhance ovarian cancer cell migration and decrease the expression of E-cadherin. Our studies also suggest that O-GlcNAcylation might become another potential target for the therapy of ovarian cancer.
Assuntos
Acetilglucosamina/metabolismo , Caderinas/metabolismo , Movimento Celular/efeitos dos fármacos , N-Acetilglucosaminiltransferases/metabolismo , Neoplasias Ovarianas/patologia , Acetilglucosamina/análogos & derivados , Acetilglucosamina/farmacologia , Antígenos CD , Western Blotting , Caderinas/genética , Cateninas/genética , Cateninas/metabolismo , Adesão Celular , Linhagem Celular Tumoral , Feminino , Inativação Gênica , Glicosilação , Humanos , Complexos Multiproteicos/genética , Complexos Multiproteicos/metabolismo , Neoplasias Ovarianas/metabolismo , Oximas/farmacologia , Fenilcarbamatos/farmacologia , Piranos/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Tiazóis/farmacologia , Transfecção , beta Catenina/genética , beta Catenina/metabolismo , delta CateninaRESUMO
According to the registered product standards specification of medical devices, combined with the standard reviewing work, the common problems of standards during medical devices registration were analyzed and corresponding suggestions were proposed to standardize the standard of the registered product, accelerate the standardization and promote the industry standardized.
Assuntos
Aprovação de Equipamentos , Equipamentos e Provisões/normasRESUMO
For the existing measurement methods of residual voltage which can't turn the power off at peak voltage exactly and simultaneously display waveforms, a new residual voltage detection method is put forward in this paper. First, the zero point of the power supply is detected with zero cross detection circuit and is inputted to a single-chip microcomputer in the form of pulse signal. Secend, when the zero point delays to the peak voltage, the single-chip microcomputer sends control signal to power off the relay. At last, the waveform of the residual voltage is displayed on a principal computer or oscilloscope. The experimental results show that the device designed in this paper can turn the power off at peak voltage and is able to accurately display the voltage waveform immediately after power off and the standard deviation of the residual voltage is less than 0.2 V at exactly one second and later.
Assuntos
Fontes de Energia Elétrica , Desenho de Equipamento , Microcomputadores , Processamento de Sinais Assistido por ComputadorRESUMO
Determiner phrases (DPs), an overarching term, can be classified into two determiner types: referential determiner phrases (RDPs, e.g., the boy) and quantificational determiner phrases (QDPs, e.g., each boy). Using the event-related potential (ERP) technique, this study explored the modulation of RDP vs. QDP in the online processing of English subject-verb agreement with omission errors by Chinese learners of English, addressing the question of whether singular quantification increases or decreases Chinese learners' sensitivity to agreement violations. The experiment manipulated the determiner type, specifically RDP vs. QDP, and grammaticality (grammatical vs. ungrammatical). The results indicated that similar to previous studies, a P600 effect was elicited in response to subject-verb agreement violations with omission errors, demonstrating that Chinese L2 learners are sensitive to such agreement violations. Additionally, the ERP patterns exhibited variations due to D-linking and number specification of RDP and QDP. Regarding D-linking, subject-verb agreement violations in the QDP conditions, necessitating integration of discourse-related knowledge, elicited laterally and frontally distributed P600 effects associated with integration complexity at the discourse level; however, non-D-linked referential determiners elicited the posteriorly-distributed P600 effects. Differences in number specification resulted in the distinctive P600 latencies and whether P600 was preceded by N400 or not. While both the RDP and QDP conditions exhibited the P600 effects, the onset latency of this effect in the number-unspecified RDP condition was 300 ms later compared to the number-specified QDP condition. Furthermore, an additional N400 component observed in the RDP condition suggests that L2 learners acquire morphologically complex subject-verb agreements by rote, treating them as unanalyzed chunks. This N400 component was absent in the QDP condition. From these results, the conclusion can be drawn that L2 learners are sensitive to the subject-verb agreement violations with omission errors, and L2 processing patterns of subject-verb agreement vary with different features of determiners, providing further evidence for the cue-based retrieval model during comprehension of grammatical sentences. Pedagogical implications are provided, and the future research direction is suggested.
RESUMO
The restoration of ancient ceramics has attracted widespread attention as it can reveal the overall appearance of ancient ceramics as well as the original information and artistic charm of cultural relics. However, traditional manual restoration is constrained due to its time-consuming nature and susceptibility to damaging ancient ceramics. Herein, a three-dimensional (3D) printing technique was employed to accurately restore Chinese Yuan Dynasty Longquan celadon using hollow Al2O3 microsphere-modified 3D printing paste. The results show that the hollow Al2O3 microsphere content plays a vital role in the printability, physical properties, and firing performance of the modified 3D printing paste. The printed green bodies show no noticeable spacing or voids under moderate rheological conditions. The as-prepared ceramic body modified with 6 wt.% hollow Al2O3 microspheres and fired at 1280 °C exhibits optimal bending strength of 56.66 MPa and a relatively low density of 2.16 gâcm-3, as well as a relatively uniform longitudinal elastic modulus and hardness along the interlayer. This 3D printing technique based on hollow Al2O3 microsphere-modified paste presents a promising pathway for achieving non-contact and damage-free restoration of cultural relics.
RESUMO
Gouty arthritis is a chronic and progressive disease characterized by high urate levels in the joints and by an inflammatory immune microenvironment. Clinical data indicate that urate reduction therapy or anti-inflammatory therapy alone often fails to deliver satisfactory outcomes. Here we have developed a smart biomimetic nanosystem featuring a 'shell' composed of a fusion membrane derived from M2 macrophages and exosomes, which encapsulates liposomes loaded with a combination of uricase, platinum-in-hyaluronan/polydopamine nanozyme and resveratrol. The nanosystem targets inflamed joints and promotes the accumulation of anti-inflammatory macrophages locally, while the uricase and the nanozyme reduce the levels of urate within the joints. Additionally, site-directed near-infrared irradiation provides localized mild thermotherapy through the action of platinum and polydopamine, initiating heat-induced tissue repair. Combined use of these components synergistically enhances overall outcomes, resulting in faster recovery of the damaged joint tissue.
RESUMO
BACKGROUND: Clear cell renal cell carcinoma (ccRCC) is a serious threat to human life. It is very important to clarify the pathogenesis of ccRCC. In this study we evaluated the clinical value of HADH and explored its role and mechanism in the malignant progression of ccRCC. METHODS: HADH expression and its relationship with prognosis were analyzed using bioinformatics database. RT-PCR, Western blot and immunohistochemistry were used to examine the expression of HADH in ccRCC tissues and tissue microarrays. To examine the cell proliferation, apoptosis, migration and invasion ability, ccRCC cells with HADH overexpressed were constructed. Xenograft experiments were performed to determine the role of HADH. Non-target metabolomics was applied to explore the potential metabolic pathway by which HADH inhibited ccRCC progression. Plasmid pcDNA3.1-NRF2 was used to confirm whether HADH inhibited the process of ccRCC cells through NRF2-related glutathione (GSH) synthesis. RESULTS: Bioinformatics database analysis showed that HADH expression was significantly decreased in ccRCC tissues, and its low expression predicted a poor prognosis. Both ccRCC tissues and tissue microarrays exhibited a significantly decreased HADH level compared with adjacent normal renal tissues. HADH overexpression inhibited the malignant behaviors of ccRCC cells. Furthermore, HADH overexpression attenuated GSH synthesis and induced oxidative stress damage. Exogenously increased NRF2 effectively attenuated the inhibitive effect of HADH overexpression on ccRCC cells. CONCLUSION: Our data revealed that HADH suppressed the malignant behaviors of ccRCC cells by attenuating GSH synthesis through inhibition of NRF2 nuclear translocation, and HADH might be a novel therapeutic target for ccRCC treatment.
RESUMO
BACKGROUND: The therapeutic potential of exosomes from human umbilical cord mesenchymal stem cells (HUMSCs-Exo) for delivering specific circular RNAs (circRNAs) in treating premature ovarian failure (POF) is not well understood. This study aimed to explore the efficacy of HUMSCs-Exo in delivering hsa_circ_0002021 for POF treatment, focusing on its effects on granulosa cell (GC) senescence and ovarian function. METHODS: Bioinformatic analysis was conducted on circRNA profiles using the GSE97193 dataset from GEO, targeting granulosa cells from varied age groups. To simulate granulosa cell senescence, KGN cells were treated with cyclophosphamide (CTX). HUMSCs were transfected with pcDNA 3.1 vectors to overexpress hsa_circ_0002021, and the HUMSCs-Exo secreted were isolated. These exosomes were characterized by transmission electron microscopy (TEM) and Western blotting to confirm exosomal markers CD9 and CD63. Co-culture of these exosomes with CTX-treated KGN cells was performed to assess ß-galactosidase activity, oxidative stress markers, ROS levels, and apoptosis via flow cytometry. Interaction between hsa_circ_0002021, microRNA-125a-5p (miR-125a-5p), and cyclin-dependent kinase 6 (CDK6) was investigated using dual-luciferase assays and RNA immunoprecipitation (RIP). A POF mouse model was induced with CTX, treated with HUMSCs-Exo, and analyzed histologically and via immunofluorescence staining. Gene expression was quantified using RT-qPCR and Western blot. RESULTS: hsa_circ_0002021 was under expressed in both in vivo and in vitro POF models and was effectively delivered by HUMSCs-Exo to KGN cells, showing a capability to reduce GC senescence. Overexpression of hsa_circ_0002021 in HUMSCs-Exo significantly enhanced these anti-senescence effects. This circRNA acts as a competitive adsorbent of miR-125a-5p, regulating CDK6 expression, which is crucial in modulating cell cycle and apoptosis. Enhanced expression of hsa_circ_0002021 in HUMSCs-Exo ameliorated GC senescence in vitro and improved ovarian function in POF models by modulating oxidative stress and cellular senescence markers. CONCLUSION: This study confirms that hsa_circ_0002021, when delivered through HUMSCs-Exo, can significantly mitigate GC senescence and restore ovarian function in POF models. These findings provide new insights into the molecular mechanisms of POF and highlight the therapeutic potential of circRNA-enriched exosomes in treating ovarian aging and dysfunction.
Assuntos
Exossomos , Células da Granulosa , Células-Tronco Mesenquimais , Insuficiência Ovariana Primária , RNA Circular , Cordão Umbilical , Insuficiência Ovariana Primária/terapia , Insuficiência Ovariana Primária/metabolismo , Insuficiência Ovariana Primária/genética , Feminino , Exossomos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Humanos , Células da Granulosa/metabolismo , RNA Circular/genética , RNA Circular/metabolismo , Animais , Cordão Umbilical/citologia , Cordão Umbilical/metabolismo , Camundongos , Senescência Celular , Apoptose , MicroRNAs/genética , MicroRNAs/metabolismoRESUMO
Introduction: Solute carrier (SLC) transport proteins play a crucial role in maintaining cellular nutrient and metabolite homeostasis and are implicated in various human diseases, making them potential targets for therapeutic interventions. However, the study of SLCs has been limited due to the lack of suitable tools, particularly cell-based substrate uptake assays, necessary for understanding their biological functions and for drug discovery purposes. Methods: In this study, a cell-based uptake assay was developed using a stable isotope-labeled compound as the substrate for SLCs, with detection facilitated by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This assay aimed to address the limitations of existing assays, such as reliance on hazardous radiolabeled substrates and limited availability of fluorescent biosensors. Results: The developed assay was successfully applied to detect substrate uptakes by two specific SLCs: L-type amino acid transporter 1 (LAT1) and sodium taurocholate co-transporting polypeptide (NTCP). Importantly, the assay demonstrated comparable results to the radioactive method, indicating its reliability and accuracy. Furthermore, the assay was utilized to screen for novel inhibitors of NTCP, leading to the identification of a potential NTCP inhibitor compound. Discussion: The findings highlight the utility of the developed cell-based uptake assay as a rapid, simple, and environmentally friendly tool for investigating SLCs' biological roles and for drug discovery purposes. This assay offers a safer alternative to traditional methods and has the potential to contribute significantly to advancing our understanding of SLC function and identifying therapeutic agents targeting SLC-mediated pathways.
RESUMO
OBJECTIVES: The clinical T1 stage solid lung cancer with metastasis is a serious threat to human life and health. In this study, we performed RNA sequencing on T1 advanced-stage lung cancer and adjacent tissues to identify a novel biomarker and explore its roles in lung cancer. METHODS: Quantitative reversed-transcription PCR, reverse transcription PCR and Western blot, MSP and Methtarget were utilized to evaluate FIBIN expression levels at both the transcriptional and protein levels as well as its methylation status. Differential target protein was evaluated for relative and absolute quantitation by isobaric tags. Co-IP was performed to detect the interactions between target protein. Precise location and expression levels of target proteins were revealed by immunofluorescence staining and component protein extraction using specific kits, respectively. RESULTS: We reported that FIBIN was frequently silenced due to promoter hypermethylation in lung cancer. Additionally, both in vitro and in vivo experiments confirmed the significant anti-proliferation and anti-metastasis capabilities of FIBIN. Mechanistically, FIBIN decreased the nuclear accumulation of ß-catenin by reducing the binding activity of GSK3ß with ANXA2 while promoting interaction between GSK3ß and ß-catenin. CONCLUSION: Our findings firstly identify FIBIN is a tumor suppressor, frequently silenced due to promoter hypermethylation. FIBIN may serve as a predictive biomarker for progression or metastasis among early-stage lung cancer patients.
Assuntos
Anexina A2 , Carcinoma Pulmonar de Células não Pequenas , Metilação de DNA , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares , Humanos , Carcinoma Pulmonar de Células não Pequenas/patologia , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Anexina A2/metabolismo , Anexina A2/genética , Animais , Camundongos , Linhagem Celular Tumoral , Proliferação de Células , beta Catenina/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Camundongos Nus , Metástase Neoplásica , Biomarcadores Tumorais/metabolismo , Biomarcadores Tumorais/genética , Masculino , Regiões Promotoras Genéticas/genética , Feminino , Camundongos Endogâmicos BALB C , Células A549 , Movimento CelularRESUMO
In our previous studies, we constructed the Bifidobacterium infantis thymidine kinase/nucleoside analogue ganciclovir (BI-TK/GCV) system, which was proven to have a sustainable antitumor activity in an in vivo bladder cancer rodent model. In this article, a proteomic approach of isobaric tags for relative and absolute quantification (iTRAQ) and followed by liquid chromatography-tandem mass spectrometry was used to understand the molecular mechanisms of this system. iTRAQ identified 192 downregulated and 210 upregulated proteins after treatment with BI-TK/GCV in Sprague-Dawley rats. Downregulations of proliferating cell nuclear antigen (PCNA), pyruvate kinase isozymes M2 (PKM2), hexokinase 1 (HXK-1), 6-phosphofructokinase (PFK-B), and cell surface glycoprotein (CD146) in bladder cancer after treatment were confirmed by Western blot analysis and validated by immunohistochemistry. Furthermore, the networks of cancer proliferation associated with PCNA, glycolysis associated with PKM2, HXK-1, and PFK-B, and invasion associated with CD146 were illustrated using Ingenuity Pathway Analysis. This study represents the successful application of iTRAQ technology to reveal the molecular mechanisms of BI-TK/GCV treatment system and provides the theoretical support for the effectiveness of our successful treatment system.
Assuntos
Bifidobacterium/metabolismo , Genes Transgênicos Suicidas , Terapia Genética , Proteômica , Simplexvirus/genética , Neoplasias da Bexiga Urinária/terapia , Animais , Bifidobacterium/genética , Western Blotting , Biologia Computacional , Ganciclovir/uso terapêutico , Regulação da Expressão Gênica/genética , Imuno-Histoquímica , Ratos , Ratos Sprague-Dawley , Neoplasias da Bexiga Urinária/genéticaRESUMO
Designed a contrast pinhole detect testing including water leak method, electrical method and improved electrical method, and concluded that the water leak method is most suitable as the arbitration method, and recommended the national standard add the requirement on electrolytic liquid filling volume of electrical test in order to improve detection accuracy.
Assuntos
Preservativos , Teste de Materiais/métodos , Látex , BorrachaRESUMO
3D-printing technology offers a flexible manufacturing platform with the potential to address the need of personalized dosage forms. However, quality aspects of such small-scale, on-demand production of pharmaceutical products intended for personalization is still limited. The aim of this study was therefore to study critical quality control attributes of lipid tablets produced by semi-solid extrusion (SSE) 3D printing from emulsion gels incorporating a poorly water-soluble drug. Quality attributes for both the printable emulsion gel and the printed dosage forms were assessed. The emulsion gel was shown to be printable with accurate dosing for at least one month of storage at 4 °C. Tablets were 3D printed in different sizes and a correlation, R2 value of 0.99, was found between the weight and the drug content. The 3D-printed tablets complied with the mass and drug content uniformity requirements described in the European Pharmacopoeia.. Solid-state characterization of the tablets during short-term storage revealed no signs of crystallinity of the drug. Lastly, the lipid digestion and drug release were unchanged after short-term storage of the tablets. This study demonstrates the potential of SSE 3D printing for personalized dosing of a lipid-based formulation strategy and discusses central quality attributes for the printable formulation and the 3D-printed dosage form.
RESUMO
INTRODUCTION: Surgery is the preferred treatment for basal cell carcinoma (BCC), locally advanced or metastatic BCC, radiation therapy or systemic therapy can be considered. Programmed death receptor 1 (PD-1) inhibitors are rarely used to treat cutaneous BCC. In the present case, we found that tislelizumab, a PD-1 immunosuppressant, had a positive effect on BCC. PATIENT CONCERNS: A 74-year-old male patient presented with a mass in the left back in October 2021, which was surgically removed and diagnosed as BCC. The patient was diagnosed with squamous lung cancer after presenting with a cough and coughing up a small amount of white, sticky sputum in December 2021. DIAGNOSIS: BCC and squamous lung cancer. INTERVENTIONS: Docetaxel + nedaplatin systemic chemotherapy combined with tislelizumab immunotherapy. OUTCOMES: Both BCC and squamous lung cancer were significantly reduced in size. CONCLUSION: After 2 cycles of immunotherapy with tislelizumab, the lung tumor shrank, the back mass disappeared, and the wound healed.
Assuntos
Carcinoma Basocelular , Carcinoma de Células Escamosas , Neoplasias Pulmonares , Neoplasias Cutâneas , Masculino , Humanos , Idoso , Neoplasias Cutâneas/complicações , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/patologia , Receptor de Morte Celular Programada 1/uso terapêutico , Carcinoma Basocelular/complicações , Carcinoma Basocelular/tratamento farmacológico , Carcinoma Basocelular/patologia , Neoplasias Pulmonares/complicações , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/patologia , Carcinoma de Células Escamosas/complicações , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/patologiaRESUMO
Excessive inflammatory reactions caused by uric acid deposition are the key factor leading to gout. However, clinical medications cannot simultaneously remove uric acid and eliminate inflammation. An M2 macrophage-erythrocyte hybrid membrane-camouflaged biomimetic nanosized liposome (USM[H]L) is engineered to deliver targeted self-cascading bienzymes and immunomodulators to reprogram the inflammatory microenvironment in gouty rats. The cell-membrane-coating endow nanosomes with good immune escape and lysosomal escape to achieve long circulation time and intracellular retention times. After being uptaken by inflammatory cells, synergistic enzyme-thermo-immunotherapies are achieved: uricase and nanozyme degraded uric acid and hydrogen peroxide, respectively; bienzymes improved the catalytic abilities of each other; nanozyme produced photothermal effects; and methotrexate has immunomodulatory and anti-inflammatory effects. The uric acid levels markedly decrease, and ankle swelling and claw curling are effectively alleviated. The levels of inflammatory cytokines and ROS decrease, while the anti-inflammatory cytokine levels increase. Proinflammatory M1 macrophages are reprogrammed to the anti-inflammatory M2 phenotype. Notably, the IgG and IgM levels in USM[H]L-treated rats decrease substantially, while uricase-treated rats show high immunogenicity. Proteomic analysis show that there are 898 downregulated and 725 upregulated differentially expressed proteins in USM[H]L-treated rats. The protein-protein interaction network indicates that the signaling pathways include the spliceosome, ribosome, purine metabolism, etc.
Assuntos
Urato Oxidase , Ácido Úrico , Ratos , Animais , Ácido Úrico/metabolismo , Ácido Úrico/farmacologia , Urato Oxidase/metabolismo , Urato Oxidase/farmacologia , Biomimética , Proteômica , Macrófagos/metabolismo , Inflamação/metabolismo , Citocinas/metabolismo , Anti-Inflamatórios/farmacologia , Membrana Eritrocítica/metabolismo , ImunoterapiaRESUMO
Dual-signal ratiometric molecularly imprinted polymer (MIP) electrochemical sensors with bimetallic active sites and high-efficiency catalytic activity were fabricated for the sensing of catechol (CC) with high selectivity and sensitivity. The amino-functionalization bimetallic organic framework materials (Fe@Ti-MOF-NH2), coupled with two-dimensional layered titanium carbide (MXene) co-modified glassy carbon electrode provides an expanded surface while amplifying the output signal through the electropolymerization immobilization of polythionine (pTHi) and MIP. The oxidation of CC and pTHi were presented as the response signal and the internal reference signal. The oxidation peak current at +0.42 V rose with increased concentration of CC, while the peak currents of pTHi at -0.20 V remained constant. Compared to the common single-signal sensing system, this one (MIP/pTHi/MXene/Fe@Ti-MOF-NH2/GCE), a novel ratiometric MIP electrochemical sensor exhibited two segments wide dynamic range of 1.0-300 µM (R2 = 0.9924) and 300-4000 µM (R2 = 0.9912), as well as an ultralow detection limit of 0.54 µM (S/N = 3). Due to the specific recognition function of MIPs and the advantages of built-in correction of pTHi, the prepared surface imprinting sensor presented an excellent performance in selectivity and reproducibility. Besides, this sensor possessed superior anti-interference ability with ions and biomolecules, excellent reproducibility, repeatability, and acceptable stability. Furthermore, the proposed sensing system exhibits high specific recognition in the determination of environmental matrices and biological fluids in real samples with satisfactory results. Therefore, this signal-enhanced ratiometric MIP electrochemical sensing strategy can accurately and selectively analyze and detect other substances.