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1.
ACS Nano ; 18(19): 12401-12411, 2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38701333

RESUMO

Accurate identification of single nucleotide variants (SNVs) in key driver genes holds a significant value for disease diagnosis and treatment. Fluorescent probes exhibit tremendous potential in specific, high-resolution, and rapid detection of SNVs. However, additional steps are required in most post-PCR assays to convert double-stranded DNA (dsDNA) products into single-stranded DNA (ssDNA), enabling them to possess hybridization activity to trigger subsequent reactions. This process not only prolongs the complexity of the experiment but also introduces the risk of losing target information. In this study, we proposed two strategies for enriching active double-stranded DNA, involving PCR based on obstructive groups and cleavable units. Building upon this, we explored the impact of modified units on the strand displacement reaction (SDR) and assessed their discriminatory efficacy for mutations. The results showed that detection of low variant allele frequencies (VAF) as low as 0.1% can be achieved. The proposed strategy allowed orthogonal identification of 45 clinical colorectal cancer tissue samples with 100% specificity, and the results were generally consistent with sequencing results. Compared to existing methods for enriching active targets, our approach offers a more diverse set of enrichment strategies, characterized by the advantage of being simple and fast and preserving original information to the maximum extent. The objective of this study is to offer an effective solution for the swift and facile acquisition of active double-stranded DNA. We anticipate that our work will facilitate the practical applications of SDR based on dsDNA.


Assuntos
DNA , Polimorfismo de Nucleotídeo Único , Polimorfismo de Nucleotídeo Único/genética , Humanos , DNA/genética , DNA/química , Neoplasias Colorretais/genética , Reação em Cadeia da Polimerase , Corantes Fluorescentes/química , DNA de Cadeia Simples/genética , DNA de Cadeia Simples/química
2.
J Nanobiotechnology ; 22(1): 189, 2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38632615

RESUMO

Although gene expression signatures offer tremendous potential in diseases diagnostic and prognostic, but massive gene expression signatures caused challenges for experimental detection and computational analysis in clinical setting. Here, we introduce a universal DNA-based molecular classifier for profiling gene expression signatures and generating immediate diagnostic outcomes. The molecular classifier begins with feature transformation, a modular and programmable strategy was used to capture relative relationships of low-concentration RNAs and convert them to general coding inputs. Then, competitive inhibition of the DNA catalytic reaction enables strict weight assignment for different inputs according to their importance, followed by summation, annihilation and reporting to accurately implement the mathematical model of the classifier. We validated the entire workflow by utilizing miRNA expression levels for the diagnosis of hepatocellular carcinoma (HCC) in clinical samples with an accuracy 85.7%. The results demonstrate the molecular classifier provides a universal solution to explore the correlation between gene expression patterns and disease diagnostics, monitoring, and prognosis, and supports personalized healthcare in primary care.


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Transcriptoma , Perfilação da Expressão Gênica , Neoplasias Hepáticas/genética , DNA , Biomarcadores Tumorais/metabolismo , Regulação Neoplásica da Expressão Gênica
3.
Inorg Chem ; 63(8): 3913-3920, 2024 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-38361417

RESUMO

Organic-inorganic hybrid perovskites (OIHPs) have received particular attention due to their characteristic structural tunability and flexibility. These features make OIHPs behave with excellent modifications on macroscopic properties, such as ferroicity or semiconductor performances, etc. Herein, we report two 2D hybrid stibium-based halide perovskite (C3H7N)3Sb2X9 (X = Br, 1; Cl, 2) ferroelastic semiconductor possessing dual switching properties of dielectric and second harmonic generation (SHG). Notably, these two hybrids exhibit halogen-regulated ferroelasticity and semiconductor properties. There is a significant difference in Curie temperature (Tc) and X-ray radiation detection sensitivity (S), i.e., the ΔTc and ΔS are 38 K and 87 µC Gyair-1 cm-2, respectively. Meanwhile, crystals 1 and 2 do not show dark current drift in cyclic measurements of different radiation doses with stable switching ratios of 30 and 10, separately. Meanwhile, these results were proven by scientific experimental results and density functional theory (DFT) calculations. Our work presents a facile and practical method to regulate macroproperties on the molecular level, providing a new vision to develop hybrid perovskite ferroic-photoelectric materials.

4.
Cancer Sci ; 115(3): 763-776, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38243657

RESUMO

Hepatocellular carcinoma (HCC) does not respond well to current treatments, even immune checkpoint inhibitors. PD-L1 (programmed cell death ligand 1 or CD274 molecule)-mediated immune escape of tumor cells may be a key factor affecting the efficacy of immune checkpoint inhibitor (ICI) therapy. However, the regulatory mechanisms of PD-L1 expression and immune escape require further exploration. Here, we observed that DDX1 (DEAD-box helicase 1) was overexpressed in HCC tissues and associated with poor prognosis in patients with HCC. Additionally, DDX1 expression correlated negatively with CD8+ T cell frequency. DDX1 overexpression significantly increased interferon gamma (IFN-γ)-mediated PD-L1 expression in HCC cell lines. DDX1 overexpression decreased IFN-γ and granzyme B production in CD8+ T cells and inhibited CD8+ T cell cytotoxic function in vitro and in vivo. In conclusion, DDX1 plays an essential role in developing the immune escape microenvironment, rendering it a potential predictor of ICI therapy efficacy in HCC.


Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Antígeno B7-H1/metabolismo , Carcinoma Hepatocelular/metabolismo , Linfócitos T CD8-Positivos , RNA Helicases DEAD-box/metabolismo , Interferon gama/metabolismo , Neoplasias Hepáticas/metabolismo , Microambiente Tumoral
5.
Sci Rep ; 13(1): 18454, 2023 10 27.
Artigo em Inglês | MEDLINE | ID: mdl-37891201

RESUMO

Clear cell renal cell carcinoma (ccRCC) is regulated by methylation modifications and long noncoding RNAs (lncRNAs). However, knowledge of N7-methylguanosine (m7G)-related lncRNAs that predict ccRCC prognosis remains insufficient. A prognostic multi-lncRNA signature was created using LASSO regression to examine the differential expression of m7G-related lncRNAs in ccRCC. Furthermore, we performed Kaplan-Meier analysis and area under the curve (AUC) analysis for diagnosis. In all, a model based on five lncRNAs was developed. Principal component analysis (PCA) indicated that the risk model precisely separated the patients into different groups. The IC50 value for drug sensitivity divided patients into two risk groups. High-risk group of patients was more susceptible to A.443654, A.770041, ABT.888, AMG.706, and AZ628. Moreover, a lower tumor mutation burden combined with low-risk scores was associated with a better prognosis of ccRCC. Quantitative real-time polymerase chain reaction (qRT-PCR) exhibited that the expression levels of LINC01507, AC093278.2 were very high in all five ccRCC cell lines, AC084876.1 was upregulated in all ccRCC cell lines except 786-O, and the levels of AL118508.1 and DUXAP8 were upregulated in the Caki-1 cell line. This risk model may be promising for the clinical prediction of prognosis and immunotherapeutic responses in patients with ccRCC.


Assuntos
Carcinoma de Células Renais , Neoplasias Renais , RNA Longo não Codificante , Humanos , Carcinoma de Células Renais/patologia , Prognóstico , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Neoplasias Renais/patologia , Estadiamento de Neoplasias , Linhagem Celular Tumoral
6.
In Vitro Cell Dev Biol Anim ; 59(9): 706-716, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37831321

RESUMO

Osteoporosis is a metabolic condition distinguished by the degradation of bone microstructure and mechanical characteristics. Traditional Chinese medicine (TCM) has been employed in China for the treatment of various illnesses. Naringin, an ingredient found in Drynariae TCM, is known to have a significant impact on bone metabolism. For this research, we studied the precise potential effect of Drynaria Naringin on protecting against bone loss caused by stress deficiency. In this study, a tail-suspension (TS) test was performed to establish a mouse model with hind leg bone loss. Some mice received subcutaneous injections of Drynaria Naringin for 30 d. Trabecular bone microarchitecture was evaluated using micro-computed tomography analysis and bone histological analysis. Bone formation and resorption markers were quantified in blood samples from mice or in the supernatant of MC3T3-E1 cells by ELISA analysis, Western blotting, and PCR. Immunofluorescence was utilized to visualize the location of ß-catenin. Additionally, siRNA was employed to knockdown-specific genes in the cells. Our findings highlight the efficacy of Drynaria Naringin in protecting against the deterioration of bone loss and promoting bone formation and Rspo1 expression in a mouse model following the TS test. Specifically, in vitro experiments also indicated that Drynaria Naringin may promote osteogenesis through the Wnt/ß-catenin signalling pathway. Moreover, our results suggest that Drynaria Naringin upregulates the expression of Rspo1/Lgr4, leading to the promotion of osteogenesis via the Wnt/ß-catenin signalling pathway. Therefore, Drynaria Naringin holds potential as a therapeutic medication for osteoporosis. Drynaria Naringin alleviates bone loss deterioration caused by mechanical stress deficiency through the Rspo1/Lgr4-mediated Wnt/ß-catenin signalling pathway.


Assuntos
Osteoporose , Polypodiaceae , Animais , Camundongos , beta Catenina/metabolismo , Diferenciação Celular , Osteogênese/genética , Osteoporose/tratamento farmacológico , Osteoporose/etiologia , Polypodiaceae/química , Estresse Mecânico , Via de Sinalização Wnt , Microtomografia por Raio-X/efeitos adversos
7.
Heliyon ; 9(6): e17078, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37484327

RESUMO

The aim of this study is to explore the active components and potential molecular mechanism of action of Rubia cordifolia L. against nasopharyngeal carcinoma (NPC). We used network pharmacology, molecular docking, and bioinformatics analysis to identify the active components and their role against NPC. The experimental verification was detected by MTT, AnnexinV-FITC/PI double fluorescence staining and Western blotting method. Network pharmacology identified that mollugin is one of the most effective components inRubia cordifolia L. Important NPC targets included HSP90AA1, CDK1, EGFR, PIK3CA, MAPK14, and CDK2. Molecular docking revealed considerable binding activity of mollugin with either of the 6 important NPC targets. Bioinformatics analysis showed that these 6 important targets were mutated in NPC, and the expression of HSP90AA1, PIK3CA, and CDK2 in cancer tissues was significantly different from that in normal tissues. MTT detection and AnnexinV-FITC/PI double fluorescence staining showed that mollugin inhibited the proliferation and induced apoptosis of NPC cells. Western blotting indicated that the molecular mechanism of mollugin against NPC was related to the regulation of the expression of Survivin and XIAP. This study predicted and partially verified the pharmacological and molecular mechanism of action of Rubia cordifolia L. against NPC. Mollugin was identified as a potential active ingredient against NPC. These results prove the reliability of network pharmacology approaches and provide a basis for further research and application of Rubia cordifolia L. against NPC.

8.
Nucleic Acids Res ; 50(20): 11727-11737, 2022 11 11.
Artigo em Inglês | MEDLINE | ID: mdl-36318259

RESUMO

CRISPR (clustered regularly interspaced short palindromic repeats) technology has achieved great breakthroughs in terms of convenience and sensitivity; it is becoming the most promising molecular tool. However, only two CRISPR activation modes (single and double stranded) are available, and they have specificity and universality bottlenecks that limit the application of CRISPR technology in high-precision molecular recognition. Herein, we proposed a novel CRISPR/Cas12a unrestricted activation mode to greatly improve its performance. The new mode totally eliminates the need for a protospacer adjacent motif and accurately activates Cas12a through toehold-mediated strand displacement and branch migration, which is highly universal and ultra-specific. With this mode, we discriminated all mismatch types and detected the EGFR T790M and L858R mutations in very low abundance. Taken together, our activation mode is deeply incorporated with DNA nanotechnology and extensively broadens the application boundaries of CRISPR technology in biomedical and molecular reaction networks.


Assuntos
Sistemas CRISPR-Cas , Neoplasias Pulmonares , Humanos , Receptores ErbB/genética , Neoplasias Pulmonares/genética , Mutação
9.
Biosens Bioelectron ; 216: 114677, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36087401

RESUMO

Nucleic acid nanotechnologies based on toehold-mediated strand displacement are ideally suited for single-nucleotide variations (SNVs) detection. But only a limited number of means could be used to construct selective hybridization probes via finely designed toehold and regulation of branching migration. Herein, we present a cooperative hybridization strategy relying on a dual-toehold and bulge-loop (DT&BL) probe, coupled with the strand displacement catalytic (SDC) cycle to identify SNVs. The dual-toehold can simultaneously hybridize the 5' and 3' ends of the target, so that it possessed the mutual correction function for improving the specificity in comparison with the single target-binding domain. Insertion of BLs into the dual-toehold probe allows tuning of Gibbs free energy change (ΔG) and control of the reaction rate during branching migration. Using the SDC cycle, the reactivity and selectivity of the DT&BL probe were increased drastically without elaborate competitive sequences. The feasibilities of this platform were demonstrated by the identification of three cancer-related genes. Moreover, the applicability of this biosensor to detect clinical samples showed satisfactory accuracy and reliability. We envision it would offer a new perspective for the construction of highly specific probes based on dynamic DNA nanotechnology, and serves as a promising tool for clinical diagnostics.


Assuntos
Técnicas Biossensoriais , Nucleotídeos , DNA/química , DNA/genética , Sondas de DNA/química , Reprodutibilidade dos Testes
10.
Front Immunol ; 13: 969447, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36032103

RESUMO

According to the difference in temperature, thermotherapy can be divided into thermal ablation and mild hyperthermia. The main advantage of thermal ablation is that it can efficiently target tumors in situ, while mild hyperthermia has a good inhibitory effect on distant metastasis. There are some similarities and differences between the two therapies with respect to inducing anti-tumor immune responses, but neither of them results in sustained systemic immunity. Malignant tumors (such as breast cancer, pancreatic cancer, nasopharyngeal carcinoma, and brain cancer) are recurrent, highly metastatic, and highly invasive even after treatment, hence a single therapy rarely resolves the clinical issues. A more effective and comprehensive treatment strategy using a combination of hyperthermia and immune checkpoint inhibitor (ICI) therapies has gained attention. This paper summarizes the relevant preclinical and clinical studies on hyperthermia combined with ICI therapies and compares the efficacy of two types of hyperthermia combined with ICIs, in order to provide a better treatment for the recurrence and metastasis of clinically malignant tumors.


Assuntos
Neoplasias da Mama , Hipertermia Induzida , Feminino , Humanos , Inibidores de Checkpoint Imunológico , Imunoterapia , Radioimunoterapia
11.
Int J Ophthalmol ; 15(5): 690-700, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35601180

RESUMO

AIM: To confirm whether exosome-mediated delivery of aptamer S58 (Exo-S58) has a better antifibrotic effect than naked S58 in human conjunctival fibroblasts (HConFs) and a rat glaucoma filtration surgery (GFS) model. METHODS: To enhance the effective reaction time of aptamer S58 in vivo, we loaded aptamer S58 into exosomes derived from HEK293T cells by PEI transfection to determine the effect of Exo-S58 in HConFs and a rat GFS model. RESULTS: Exo-S58 can significantly reduce cell proliferation, migration and fibrosis in TGF-ß2-induced HConFs. In an in vivo experiment, Exo-S58 treatment prolonged filtering bleb retention and reduced fibrosis compared with naked S58 treatment in GFS rats. CONCLUSION: The exosomes are safe and valid carriers to deliver aptamers. Furthermore, Exo-S58 exhibited superior antifibrotic effect than naked S58 both in HConFs cells and rat GFS models.

12.
J Healthc Eng ; 2022: 6277139, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35463684

RESUMO

Background: Nasopharyngeal carcinoma (NPC), a neoplasm of the head and neck, has high incidence and mortality rates in East and Southeast Asia. Evodia rutaecarpa is a tree native to Korea and China, and its fruit (hereafter referred to as Evodia) exhibits remarkable antitumour properties. However, little is known about its mechanism of action in NPC. In this study, we employed network pharmacology to identify targets of active Evodia compounds in nasopharyngeal carcinoma and generate an interaction network. Methods: The active ingredients of Evodia and targets in NPC were obtained from multiple databases, and an interaction network was constructed via the Cytoscape and STRING databases. The key biological processes and signalling pathways were predicted using Gene Ontology and Kyoto Encyclopaedia of Genes and Genomes pathway enrichment analyses. Molecular docking technology was used to identify the affinity and activity of target genes, and The Cancer Genome Atlas and Human Protein Atlas databases were used to analyse differential expression. Cell Counting Kit-8 (CCK-8) and Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) dual-fluorescence staining were used for experimental verification. Results: Active Evodia compounds included quercetin, isorhamnetin, and evodiamine, and important NPC targets included MAPK14, AKT1, RELA, MAPK1, JUN, and p53, which were enriched in lipid and atherosclerosis signalling pathways. Additionally, we verified the high affinity and activity of the active compounds through molecular docking, and the target proteins were verified using immunohistochemistry and differential expression analyses. Furthermore, CCK-8 assays and Annexin V-FITC/PI dual-fluorescence staining showed that isorhamnetin inhibited the proliferation of NPC cells and induced apoptosis. Conclusion: Our results identified the molecular mechanisms of Evodia and demonstrated its ability to alter the proliferation and apoptosis of NPC cells through multiple targets and pathways, thereby providing evidence for the clinical application of Evodia.


Assuntos
Medicamentos de Ervas Chinesas , Evodia , Neoplasias Nasofaríngeas , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Frutas , Humanos , Medicina Tradicional Chinesa/métodos , Simulação de Acoplamento Molecular , Carcinoma Nasofaríngeo/tratamento farmacológico , Neoplasias Nasofaríngeas/tratamento farmacológico , Farmacologia em Rede
13.
Bioengineered ; 13(3): 6688-6697, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35235756

RESUMO

Oxaliplatin is an effective chemotherapeutic agent for the treatment of malignant tumors. However, severe oxaliplatin-induced neurotoxicity has been well documented. Memantine is a drug for the management of Alzheimer's Disease (AD) due to its promising neuroprotective properties. We hypothesize that Memantine possesses a beneficial role against chemotherapy-induced neuronal damages. In this study, we established an oxaliplatin-induced neurotoxicity assay model in human SHSY-5Y neuronal cells and investigated the protective effect of Memantine. We showed that Memantine treatment ameliorated oxaliplatin-elevated intracellular production of reactive oxygen species (ROS), lipid product malondialdehyde (MDA), and NOX-2 expression. Memantine alleviated impairment of the mitochondrial membrane potential and ATP production by oxaliplatin. As a result, Memantine showed a protective role against oxaliplatin-induced cytotoxicity. Moreover, the terminal deoxynucleotidyl Transferase-mediated dUTP nick end labeling (TUNEL) apoptosis assay revealed that Memantine protected oxaliplatin-induced apoptosis through mitigating the ratio of Bax/Bcl-2 and Caspase-3 cleavage. We concluded Memantine ameliorated the neurotoxicity of oxaliplatin in a mitochondrial-dependent pathway.


Assuntos
Memantina , Mitocôndrias , Apoptose , Humanos , Memantina/metabolismo , Memantina/farmacologia , Potencial da Membrana Mitocondrial , Mitocôndrias/metabolismo , Oxaliplatina/farmacologia , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo
14.
Comb Chem High Throughput Screen ; 25(12): 2070-2081, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35048799

RESUMO

BACKGROUND: Cancer stem cells (CSCs) are now being considered as the initial component in the development of pancreatic adenocarcinoma (PAAD). Our aim was to develop a CSCrelated signature to assess the prognosis of PAAD patients for the optimization of treatment. METHODS: Differentially expressed genes (DEGs) between pancreatic tumor and normal tissue in the Cancer Genome Atlas (TCGA) were screened out, and the weighted gene correlation network analysis (WGCNA) was employed to identify the CSC-related gene sets. Then, univariate, Lasso Cox regression analyses and multivariate Cox regression were applied to construct a prognostic signature using the CSC-related genes. Its prognostic performance was validated in TCGA and ICGC cohorts. Furthermore, Univariate and multivariate Cox regression analyses were used to identify independent prognostic factors in PAAD, and a prognostic nomogram was established. RESULTS: The Kaplan-Meier analysis, ROC curve and C-index indicated the good performance of the CSC-related signature at predicting overall survival (OS). Univariate Cox regression and multivariate Cox regression revealed that the CSC-related signature was an independent prognostic factor in PAAD. The nomogram was superior to the risk model and AJCC stage in predicting OS. In terms of mutation and tumor immunity, patients in the high-risk group had higher tumor mutation burden (TMB) scores than patients in the low-risk group, and the immune score and the ESTIMATE score were significantly lower in the high-risk group. Moreover, according to the results of principal component analysis (PCA) and Gene Set Enrichment Analysis (GSEA), the low-risk and high-risk groups displayed different stemness statuses based on the risk model. CONCLUSION: Our study identified four CSC-related gene signatures and established a prognostic nomogram that reliably predicts OS in PAAD. The findings may support new ideas for screening therapeutic targets to inhibit stem characteristics and the development of PAAD.


Assuntos
Adenocarcinoma , Neoplasias Pancreáticas , Adenocarcinoma/diagnóstico , Adenocarcinoma/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Células-Tronco Neoplásicas/patologia , Nomogramas , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/genética , Prognóstico , Neoplasias Pancreáticas
15.
Ann Surg Oncol ; 29(3): 2034-2040, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34647203

RESUMO

BACKGROUND: Laparoscopic anatomic mesohepatectomy for patients with hepatocellular carcinoma (HCC) remains technically challenging, especially for those with a massive tumor larger than 10 cm. METHODS: In this study, a 65-year-old man with a 13 × 10-cm2 solitary liver tumor located at segments 4, 5, and 8 underwent laparoscopic mesohepatectomy. To reduce the possibility of releasing cancer cells from the primary tumor, the in situ resection strategy for tumor removal was implemented. The intrafascial approach was used to dissect the right Glissonean pedicle, to transect the right anterior hepatic artery, and to ligate the right anterior portal vein. The extrafascial and transfissural approach was performed along the umbilical fissure to transect the Glissonean pedicle of segment 4. Indocyanine green (ICG) then was applied using "reverse staining" to visualize the resection extent and the right posterior hepatic duct (RPHD). During parenchymal resection, the right anterior Glissonean pedicle was adequately exposed and transected via the extrafascial approach above the plane of the RPHD. Finally, the right coronary ligament was dissected, and the tumor was removed. RESULTS: The operation was completed in 360 min, with a blood loss of 200 mL. The histopathologic diagnosis indicated a moderately differentiated HCC. The patient was discharged on postoperative day 8 without any complications. CONCLUSION: Laparoscopic in situ anatomic mesohepatectomy using combined intra- and extrafascial approaches with ICG navigation may be feasible for patients with a centrally located solitary massive HCC.


Assuntos
Carcinoma Hepatocelular , Laparoscopia , Neoplasias Hepáticas , Idoso , Carcinoma Hepatocelular/cirurgia , Hepatectomia , Humanos , Verde de Indocianina , Neoplasias Hepáticas/cirurgia , Masculino
16.
Biosens Bioelectron ; 194: 113625, 2021 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-34534950

RESUMO

N6-methyladenosine (m6A) is the most abundant post-transcriptional modification in RNA and has important implications in physiological processes and tumor development. However, sensitive and specific quantification of locus-specific m6A modification levels remains a challenging task. In the present work, a novel m6A-sensitive DNAzyme was utilized to directly detect m6A by coupling with a three-way junction-mediated isothermal exponential CRISPR amplification reaction for the first time. This method was built on the fact that the binding arm of the DNAzyme bound to the specific site and its core structure catalyzed the selective cleavage of unmodified adenine instead of methylated adenines. Subsequently, the intact RNA was identified by the proximity effect of the three-way junction. Enormous amounts of single-stranded DNA products were generated through a combination of SDA and EXPAR for signal amplification. The specific real-time curve of products was recorded through detecting the fluorescence intensity triggered by CRISPR Cas12a. As a result, methylation target of abundance down to 1% was successfully identified. In addition, this strategy could be used for the analysis of cell RNA extracts. Combined with an electrochemical sensor for quantitative detection of RNA methylation, we demonstrated the generality of as-proposed strategy. We envision the present method would provide a new platform for the analysis of m6A in RNA and promote its application in clinical diseases.


Assuntos
Técnicas Biossensoriais , DNA Catalítico , Adenosina/análogos & derivados , Metilação , RNA/metabolismo
17.
Aging (Albany NY) ; 13(13): 17607-17628, 2021 07 08.
Artigo em Inglês | MEDLINE | ID: mdl-34237706

RESUMO

The function of competitive endogenous RNA (ceRNA) network in the immune regulation of hepatocellular carcinoma (HCC) is unclear. Our study aimed to construct an immune-related ceRNA network and develop an immune-related long noncoding RNA (lncRNA) signature to assess the prognosis of HCC patients and to optimize the treatment methods. We firstly constructed a ceRNA regulatory network for HCC using differentially expressed lncRNAs, mRNAs and microRNAs (miRNAs) from the Cancer Genome Atlas. A signature was constructed by 11 immune-related prognostic lncRNAs from the ceRNA network. The survival analysis and receiver operating characteristic analysis validated the reliability of the signature. Multivariate Cox regression analysis revealed that the signature could act an independent prognostic indicator. This signature also showed high association with immune cell infiltration and immune check blockades. LINC00491 was identified as the hub lncRNA in the signature. In vitro and in vivo evidence demonstrated that silencing of LINC00491 significantly inhibited HCC growth. Finally, 59 lncRNAs, 21 miRNAs, and 26 mRNAs were obtained to build the immune-related ceRNA network for HCC. In conclusion, our novel immune-related lncRNA prognostic signature and the immune-related ceRNA network might provide in-depth insights into tumor-immune interaction of HCC and promote better individual treatment strategies in HCC patients.


Assuntos
Biomarcadores Tumorais/análise , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/genética , RNA Longo não Codificante/análise , RNA Longo não Codificante/genética , Animais , Carcinoma Hepatocelular/imunologia , Redes Reguladoras de Genes/genética , Inativação Gênica , Genômica , Humanos , Neoplasias Hepáticas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Prognóstico , Curva ROC , Reprodutibilidade dos Testes , Análise de Sobrevida , Ensaios Antitumorais Modelo de Xenoenxerto
18.
Surg Oncol ; 38: 101622, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34146767

RESUMO

INTRODUCTION: Segment 8 is considered the largest liver segment, and its portal vein branches are generally divided into four parts, including ventral, dorsal, dorsolateral and medial branches (Shindoh et al., 2010; Takayasu et al., 1985) [1,2]. An anatomic combined subsegmentectomy could satisfy both the oncological quality of anatomical resection and the safety of parenchyma sparing principle if a small hepatocellular carcinoma is located between the hepatic subsegments (Berardi et al., 2021) [3]. Yet, laparoscopic anatomic combined subsegmentectomy of segment 8 is still technically challenging. The development of digital intelligent technology has made it possible to tailored preoperative planning and accurate intraoperative navigation in laparoscopic surgery. VIDEO: A 57-year-old man underwent a routine CT scan and was found to have a mass occupation in segment 8 of the liver. Three-dimensional reconstruction was performed to evaluate liver anatomy, vascular variations, and volume of each vascular unit as well as the location of the tumor, its relationship with the liver anatomy, and the Glissonian pedicles feeding the tumor-bearing area. Based on the reconstructed model, resection was planned aiming to the narrowest but oncologically safe anatomical tumor-bearing area. Upon evaluation, anatomic combined subsegmentectomy of segment 8 (ventral and medial subsegments) was confirmed. The operation was performed precisely under assistance of the Laparoscopic Hepatectomy Navigation System (LHNS, software copyright No. 2018SR840555) (Yang et al., 2020) [4]. RESULTS: The operation lasted 200 min with 50 ml intraoperative blood loss. There were no postoperative complications, and the patient was discharged after 6 days. CONCLUSION: Digital intelligent technology could provide tailored strategy for laparoscopic liver surgery, which makes laparoscopic anatomic combined subsegmentectomy of segment 8 feasible and effective.


Assuntos
Veias Hepáticas/cirurgia , Laparoscopia/métodos , Neoplasias Hepáticas/cirurgia , Veia Porta/cirurgia , Tomografia Computadorizada por Raios X/métodos , Veias Hepáticas/diagnóstico por imagem , Veias Hepáticas/patologia , Humanos , Neoplasias Hepáticas/diagnóstico por imagem , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Veia Porta/diagnóstico por imagem , Veia Porta/patologia , Prognóstico , Gravação em Vídeo
19.
Am J Transl Res ; 13(4): 2983-2989, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34017465

RESUMO

OBJECTIVE: To investigate the effects of nursing based on Orem's self-care model on self-care efficacy, quality of life (QOL) and adverse emotions of patients with advanced lung cancer (ALC) receiving chemotherapy. METHODS: A total of 71 patients with ALC aged 50-70 years, from our hospital were selected as the study subjects and divided into the control group (CNG, n = 35) and the experimental group (EXG, n = 36) using the random number table method. The CNG was treated with conventional chemotherapy combined with conventional nursing, while the EXG was treated with conventional chemotherapy combined with nursing based on Orem's self-care model. The effects on self-care efficacy, QOL and adverse emotions in the two groups were observed before and after nursing. The General Self-Efficacy Scale (GSES) was scored in both groups. The patients' body, physiology, psychology, society and health were scored using the QOL questionnaire for Chinese cancer patients receiving chemotherapy (QLQ-CCC). The self-rating anxiety scale (SAS) and self-rating depression scale (SDS) in the two groups were scored using the Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD). RESULTS: The GSES scores in the EXG were remarkably higher than those in the CNG after intervention (P < 0.05). After intervention, the scores of the patients' body, physiology, psychology, society and health in the EXG were higher than those in the CNG (P < 0.05). The scores of SAS and SDS in the EXG were lower than those in the CNG (P < 0.05). CONCLUSION: Nursing based on Orem's self-care model can effectively improve the self-care efficacy and QOL, adverse emotions (e.g., anxiety and depression), and degree of pain of patients with ALC receiving chemotherapy. Therefore, it has a positive clinical significance.

20.
World J Surg ; 45(7): 2108-2115, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33770240

RESUMO

OBJECTIVES: To evaluate the clinical impact and technical feasibility of augmented reality laparoscopic navigation (ARLN) system in laparoscopic splenectomy for massive splenomegaly. METHODS: The clinical data of 17 consecutive patients who underwent laparoscopic splenectomy using ARLN (ARLN group) and 26 patients without ARLN guidance (Non-ARLN group) between January 2018 and April 2020 were enrolled. Propensity score matching (PSM) analysis was performed between the patients with and without ARLN guidance at a ratio of 1:1. RESULTS: Mean intraoperative blood loss was significantly lower in the ARLN-group than in the Non-ARLN group (306.6 ml vs. 462.6 ml, p = 0.047). All the patients in the ARLN-group achieved successful splenic artery dissection, while surgical success was achieved in 12 patients in the Non-ARLN group (p = 0.044). Postoperative hospital stay was significantly longer in the Non-ARLN group (3.8 days vs. 4.5 days, p = 0.040). CONCLUSIONS: ARLN can provide feasible and accurate intraoperative image guidance, and it could be helpful in the performance of laparoscopic splenectomy for massive splenomegaly.


Assuntos
Realidade Aumentada , Laparoscopia , Humanos , Estudos Retrospectivos , Esplenectomia , Esplenomegalia/cirurgia , Resultado do Tratamento
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