RESUMO
BACKGROUND AND AIMS: The innate-like mucosa-associated invariant T (MAIT) cells are enriched in human liver and have been linked to human HCC. However, their contributions to the progression of HCC are controversial due to the heterogeneity of MAIT cells, and new MAIT cell subsets remain to be explored. APPROACH AND RESULTS: Combining single cell RNA sequencing (scRNA-seq) and flow cytometry analysis, we performed phenotypic and functional studies and found that FOXP3 + CXCR3 + MAIT cells in HCC patients were regulatory MAIT cells (MAITregs) with high immunosuppressive potential. These MAITregs were induced under Treg-inducing condition and predominantly from FOXP3 - CXCR3 + MAIT cells, which displayed mild Treg-related features and represented a pre-MAITreg reservoir. In addition, the induction and function of MAITregs were promoted by ß1 adrenergic receptor signaling in pre-MAITregs and MAITregs, respectively. In HCC patients, high proportion of the intratumoral MAITregs inhibited antitumor immune responses and was associated with poor clinical outcomes. CONCLUSIONS: Together, we reveal an immunosuppressive subset of MAIT cells in HCC patients that contributes to HCC progression, and propose a control through neuroimmune crosstalk.
Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Células T Invariantes Associadas à Mucosa , Humanos , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Mucosa , Fatores de Transcrição Forkhead , Receptores AdrenérgicosRESUMO
Adaptive optics using direct wavefront sensing (direct AO) is widely used in two-photon microscopy to correct sample-induced aberrations and restore diffraction-limited performance at high speeds. In general, the direct AO method employs a Sharked-Hartman wavefront sensor (SHWS) to directly measure the aberrations through a spot array. However, the signal-to-noise ratio (SNR) of spots in SHWS varies significantly within deep tissues, presenting challenges for accurately locating spot centroids over a large SNR range, particularly under extremely low SNR conditions. To address this issue, we propose a piecewise centroid calculation algorithm called GCP, which integrates three optimal algorithms for accurate spot centroid calculations under high-, medium-, and low-SNR conditions. Simulations and experiments demonstrate that the GCP can accurately measure aberrations over a large SNR range and exhibits robustness under extremely low-SNR conditions. Importantly, GCP improves the AO working depth by 150â µm compared to the conventional algorithm.
RESUMO
Compared to intensity detection, fluorescence lifetime has the advantage of being unaffected by variations in excitation intensity, fluorophore concentration, or attenuation due to biological absorption and scattering. In this Letter, to the best of our knowledge, we present the use of the two-photon excitation autofluorescence lifetime imaging of tryptophan (TRP) to probe cell metabolism for the first time. Tests of pure chemical samples showed that the fluorescence lifetime of TRP was highly sensitive to changes in molecular conformation and the environment. In in vitro cell experiments, we successfully utilized the fluorescence lifetime of TRP to distinguish tumor cells from healthy cells, track the therapeutic effect of the tumor immunotherapy drug 1-MT for HeLa cells, and monitor cells in response to carbonyl cyanide 3-chlorophenylhydrazone (CCCP)-induced cell apoptosis. These results reveal that the two-photon excitation autofluorescence lifetime of TRP could be a sensitive natural probe of cell metabolism in living cells.
Assuntos
Triptofano , Humanos , Células HeLa , Triptofano/químicaRESUMO
Severe fever with thrombocytopenia syndrome (SFTS), which is caused by a novel Bunyavirus, has gradually become a threatening infectious disease in rural areas of Asia. Studies have identified a severe cytokine storm and impaired humoral immune response in SFTS. However, the cellular immune response to SFTS virus (SFTSV) infection remains largely unknown. Here we report that SFTS patients had a cytokine storm accompanied by high levels of chemokines. CD8+ T cells in peripheral blood mononuclear cells of SFTS patients exhibited a more activated phenotype and enhanced the antiviral responses. They increased the expression of CD69 and CD25, secreted a higher level of IFN-γ and granzyme, and had a stronger proliferative ability than in healthy controls. In convalescent SFTS patients, the expression of CD69 and CD25 on CD8+ T cells was reduced. In addition, we found the ratio and cellularity of CD14+ CD16+ intermediate monocytes were increased in peripheral blood of SFTS patients. Both the expression of C-X-C motif chemokine ligand 10 (CXCL10) on CD14+ CD16+ intermediate monocytes and the expression of C-X-C motif chemokine receptor 3 (CXCR3) on CD8+ T cells increased dramatically in SFTS patients. Our studies reveal a potential pathway that CD8+ T cells rapidly activate and are mostly recruited by intermediate monocytes through CXCL10 in SFTSV infection. Our results may be of clinical relevance for further treatment and discharge instructions in SFTSV infections.
Assuntos
Infecções por Bunyaviridae , Phlebovirus , Febre Grave com Síndrome de Trombocitopenia , Trombocitopenia , Humanos , Febre Grave com Síndrome de Trombocitopenia/tratamento farmacológico , Infecções por Bunyaviridae/tratamento farmacológico , Linfócitos T CD8-Positivos , Leucócitos Mononucleares , Síndrome da Liberação de Citocina , Trombocitopenia/tratamento farmacológico , Antivirais/uso terapêuticoRESUMO
BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is a prevalent condition that often goes unrecognized in the population, and many risk factors for this disease are not well understood. Glyphosate (GLY) is one of the most commonly used herbicides worldwide, and exposure to this chemical in the environment is significant. However, studies exploring the association between GLY exposure and NAFLD remain limited. Therefore, the aim of this study was to assess the association between urinary glyphosate (uGLY) level and fatty liver index (FLI) using data from the National Health and Nutrition Examination Survey (NHANES), which includes uGLY measurements. METHODS: The log function of uGLY was converted and expressed as Loge(uGLY) with the constant "e" as the base and used for subsequent analysis. The association between Loge(uGLY) (the independent variable) level and FLI (the dependent variable) was assessed by multiple linear regression analysis. Smoothing curve fitting and a generalized additive model were used to assess if there was a nonlinear association between the independent and the dependent variables. A subgroup analysis was used to find susceptible individuals of the association between the independent variable and the dependent variable. RESULTS: A final total of 2238 participants were included in this study. Participants were categorized into two groups (< -1.011 and ≥ -1.011 ng/ml) based on the median value of Loge(uGLY). A total of 1125 participants had Loge(uGLY) levels ≥ -1.011 ng/ml and higher FLI. The result of multiple linear regression analysis showed a positive association between Loge(uGLY) and FLI (Beta coefficient = 2.16, 95% CI: 0.71, 3.61). Smoothing curve fitting and threshold effect analysis indicated a linear association between Loge(uGLY) and FLI [likelihood ratio(LLR) = 0.364]. Subgroup analyses showed that the positive association between Loge(uGLY) and FLI was more pronounced in participants who were female, aged between 40 and 60 years, had borderline diabetes history, and without hypertension history. In addition, participants of races/ethnicities other than (Mexican American, White and Black) were particularly sensitive to the positive association between Loge(uGLY) and FLI. CONCLUSIONS: A positive linear association was found between Loge(uGLY) level and FLI. Participants who were female, 40 to 60 years old, and of ethnic backgrounds other than Mexican American, White, and Black, deserve more attention.
Assuntos
Hipertensão , Hepatopatia Gordurosa não Alcoólica , Adulto , Humanos , Feminino , Pessoa de Meia-Idade , Masculino , Glifosato , Hepatopatia Gordurosa não Alcoólica/epidemiologia , Inquéritos Nutricionais , EtnicidadeRESUMO
PURPOSE: To develop predictive nomograms of lymph vascular space invasion (LVSI) in patients with early-stage cervical cancer. METHODS: We identified 403 patients with cervical cancer from the Affiliated Hospital of Jiangnan University from January 2015 to December 2019. Patients were divided into the training set (n = 242) and the validation set (n = 161), with patients in the training set subdivided into LVSI (+) and LVSI (-) groups according to postoperative pathology. Preoperative hematologic indexes were compared between the two subgroups. Univariate and multivariate logistic regression analyses were used to analyze the independent risk factors for LVSI, from which a nomogram was constructed using the R package. RESULTS: LVSI (+) was present in 94 out of 242 patients in the training set, accompanied by a significant increase in the preoperative squamous cell carcinoma antigen (SCC), white blood cells (WBC), neutrophil (NE), platelet (PLT), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), systemic inflammation index (SII), and tumor size (P < 0.05). Univariate analysis showed that SCC, WBC, NE, NLR, PLR, SII, and tumor size were correlated with LVSI (P < 0.05), and multivariate analysis showed that tumor size, SCC, WBC, and NLR were independent risk factors for LVSI (P < 0.05). A nomogram was correspondingly established with good performance in predicting LVSI [training: ROC-AUC = 0.845 (95% CI: 0.731-0.843) and external validation: ROC-AUC = 0.704 (95% CI: 0.683-0.835)] and high accuracy (training: C-index = 0.787; external validation: C-index = 0.759). CONCLUSION: The nomogram based on preoperative tumor size, SCC, WBC, and NLR had excellent accuracy and discriminative capability to assess the risk of LVSI in early-stage cervical cancer patients.
Assuntos
Nomogramas , Neoplasias do Colo do Útero , Feminino , Humanos , Neoplasias do Colo do Útero/cirurgia , Neoplasias do Colo do Útero/patologia , Estudos Retrospectivos , Colo do Útero/patologia , Fatores de Risco , InflamaçãoRESUMO
OBJECTIVE: Haematogenous dissemination is a prevalent route of colorectal cancer (CRC) metastasis. However, as the gatekeeper of vessels, the role of tumour pericytes (TPCs) in haematogenous metastasis remains largely unknown. Here, we aimed to investigate the heterogeneity of TPCs and their effects on CRC metastasis. DESIGN: TPCs were isolated from patients with CRC with or without liver metastases and analysed by single-cell RNA sequencing (scRNA-seq). Clinical CRC specimens were collected to analyse the association between the molecular profiling of TPCs and CRC metastasis. RNA-sequencing, chromatin immunoprecipitation-sequencing and bisulfite-sequencing were performed to investigate the TCF21-regulated genes and mechanisms underlying integrin α5 on TCF21 DNA hypermethylation. Pericyte-conditional Tcf21-knockout mice were constructed to investigate the effects of TCF21 in TPCs on CRC metastasis. Masson staining, atomic force microscopy, second-harmonic generation and two-photon fluorescence microscopy were employed to observe perivascular extracellular matrix (ECM) remodelling. RESULTS: Thirteen TPC subpopulations were identified by scRNA-seq. A novel subset of TCF21high TPCs, termed 'matrix-pericytes', was associated with liver metastasis in patients with CRC. TCF21 in TPCs increased perivascular ECM stiffness, collagen rearrangement and basement membrane degradation, establishing a perivascular metastatic microenvironment to instigate colorectal cancer liver metastasis (CRCLM). Tcf21 depletion in TPCs mitigated perivascular ECM remodelling and CRCLM, whereas the coinjection of TCF21high TPCs and CRC cells markedly promoted CRCLM. Mechanistically, loss of integrin α5 inhibited the FAK/PI3K/AKT/DNMT1 axis to impair TCF21 DNA hypermethylation in TCF21high TPCs. CONCLUSION: This study uncovers a previously unidentified role of TPCs in haematogenous metastasis and provides a potential diagnostic marker and therapeutic target for CRC metastasis.
Assuntos
Neoplasias Colorretais , Neoplasias Hepáticas , Animais , Camundongos , Linhagem Celular Tumoral , Neoplasias Colorretais/patologia , DNA , Regulação Neoplásica da Expressão Gênica , Integrina alfa5/genética , Integrina alfa5/metabolismo , Neoplasias Hepáticas/patologia , Metástase Neoplásica , Pericitos/metabolismo , Pericitos/patologia , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Microambiente TumoralRESUMO
As the key component of host innate antiviral immunity, type I interferons (IFN-Is) exert multiple antiviral effects by inducing hundreds of IFN-stimulated genes. However, the precise mechanism involved in host sensing of IFN-I signaling priming is particularly complex and remains incompletely resolved. This research identified F-box protein 11 (FBXO11), a component of the E3-ubiquitin ligase SKP/Cullin/F-box complex, acted as an important regulator of IFN-I signaling priming and antiviral process against several RNA/DNA viruses. FBXO11 functioned as an essential enhancer of IFN-I signaling by promoting the phosphorylation of TBK1 and IRF3. Mechanistically, FBXO11 facilitated the assembly of TRAF3-TBK1-IRF3 complex by mediating the K63 ubiquitination of TRAF3 in a NEDD8-dependent manner to amplify the activation of IFN-I signaling. Consistently, the NEDD8-activating enzyme inhibitor MLN4921 could act as a blocker for FBXO11-TRAF3-IFN-I axis of signaling. More significantly, examination of clinical samples of chronic hepatitis B virus (HBV) infection and public transcriptome database of severe acute respiratory syndrome coronavirus-2-, HBV-, and hepatitis C virus-infected human samples revealed that FBXO11 expression was positively correlated with the stage of disease course. Taken together, these findings suggest that FBXO11 is an amplifier of antiviral immune responses and might serve as a potential therapeutic target for a number of different viral diseases.
Assuntos
COVID-19 , Proteínas F-Box , Hepatite B Crônica , Interferon Tipo I , Humanos , Antivirais/farmacologia , Proteínas Serina-Treonina Quinases/genética , Fator 3 Associado a Receptor de TNF/genética , Imunidade Inata , Interferon Tipo I/metabolismo , Fator Regulador 3 de Interferon/genética , Proteína-Arginina N-Metiltransferases/metabolismoRESUMO
Two-photon microscopy (TPM) has provided critical in situ and in vivo information in biomedical studies due to its high resolution, intrinsic optical sectioning, and deep penetration. However, its relatively small field of view (FOV), which is usually determined by objectives, restricts its wide application. In this paper, we propose a segment-scanning sensorless adaptive optics method to extend the FOV and achieve high-resolution and large-FOV two-photon imaging. We demonstrated the proposed method by imaging fluorescent beads, cerebral nerve cells of mouse brain slices, and cerebral vasculature and microglia of live mice. The method extended the FOV of a commercial objective from 1.8 to 3.46 mm while maintaining a lateral resolution of 840 nm and high signal-to-noise ratio. Our technology is compatible with a standard TPM and can be used for large-scale biological exploration.
Assuntos
Microscopia , Fótons , Animais , Camundongos , Óptica e Fotônica , Razão Sinal-RuídoRESUMO
As the pesticide and its metabolite residues in processed fruits could become a significant route of human exposure. The work presented herein focuses on developing a feasible quick, easy, cheap, effective, rugged, and safe method with improved extraction and cleanup system for the determination of imidacloprid, acetamiprid, thiamethoxam, and clothianidin (a metabolite of thiamethoxam) in canned fruits. The low toxic solvent ethanol was used to extract the analytes, and ammonium sulfate was used to promote phase separation. Moreover, the carboxylated multi-walled carbon nanotube acted as the clean-up sorbent for the removal of high solubility impurities. The proposed method was validated with fortified real samples at different concentration levels (20-200 µg/kg). Recoveries obtained from three spiked levels (20, 50, and 200 µg/kg) ranged from 74.9 to 86.4% with relative standard deviations of the intra-day and inter-day in the range of 0.8-5.5 and 2.0-7.1%, respectively. The limits of detection ranged from 0.2 to 0.5 and 0.2 to 0.6 µg/kg for orange and peach, respectively. The results demonstrated that the proposed method could be considered appropriate, and comparatively lower toxic for the analysis of neonicotinoid pesticide residues in canned fruit.
Assuntos
Resíduos de Praguicidas , Praguicidas , Sulfato de Amônio , Etanol , Frutas/química , Humanos , Neonicotinoides/análise , Resíduos de Praguicidas/análise , Praguicidas/análise , Extração em Fase Sólida/métodos , Tiametoxam/análiseRESUMO
Compressive sensing (CS) is a signal sampling theory that originated about 16 years ago. It replaces expensive and complex receiving devices with well-designed signal recovery algorithms, thus simplifying the imaging system. Based on the application of CS theory, a single-pixel camera with an array-detection imaging system is established for high-pixel detection. Each detector of the detector array is coupled with a bundle of fibers formed by fusion of four bundles of fibers of different lengths, so that the target area corresponding to one detector is split into four groups of target information arriving at different times. By comparing the total amount of information received by the detector with the threshold set in advance, it can be determined whether the four groups of information are calculated separately. The simulation results show that this new system can not only reduce the number of measurements required to reconstruct high quality images but can also handle situations wherever the target may appear in the field of view without necessitating an increase in the number of detectors.
RESUMO
BACKGROUND: The development of rapid and sensitive monitoring methods for trace N-nitrosamines (NAs) in foodstuffs is essential for mitigating the potential health risks to consumers. In the present study, an analytical platform based on one step fully automated static headspace sampling and gas chromatography-tandem mass spectrometry (GC-MS/MS) was developed and validated for the analysis of N-nitrosamines in dried aquatic products of animal origin. The samples and sodium chloride solution mixture were incubated in a heated headspace vial for analyte evaporation, coupled to automatic sampling and online GC-MS/MS analysis. The proposed method requires minimal sample preparation and organic solvent consumption. Five N-nitrosamines including N-nitroso dimethylamine, N-nitroso methyl ethylamine, N-nitroso pyrolidine, N-nitroso piperidine and N-nitroso diphenylamine were selected as model compounds to optimize the significant factors by a using Box-Behnken design. RESULTS: The optimum conditions achieved limits of detections in the range 0.08-0.29 µg kg-1 , with correlation coefficient over 0.998. Relative recoveries in dried aquatic product sample were in the range 76.9-92.4%, with relative SDs of 1.9-7.2%. CONCLUSION: These results confirm the reliability of the developed method for further application in trace level monitoring of the target analytes in foodstuffs. © 2022 Society of Chemical Industry.
Assuntos
Nitrosaminas , Animais , Nitrosaminas/análise , Nitrosaminas/química , Espectrometria de Massas em Tandem/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Reprodutibilidade dos Testes , Dimetilnitrosamina/análiseRESUMO
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is sweeping the world since the end of 2019. The titer change of antibodies against SARS-CoV-2 needs to be further clarified, the clinical and preventive value of antibodies still needs to be further investigated. An enzyme-linked immunosorbent assay (ELISA) was established by coating with SARS-CoV-2 recombinant spike protein and used to detect serum immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies against SARS-CoV-2 in coronavirus disease 2019 patients to evaluate the pattern of changes of antibodies. The specificity of the ELISA for detection SARS-CoV-2 IgM and IgG were 96% (144/150) and 100% (150/150), respectively. The sensitivity of ELISA was 100% (150/150) for IgM, and 99.3% (149/150) for IgG. SARS-CoV-2-SP-IgM and SP-IgG antibodies could be detected on Day 1 of hospitalization in 12.5% patients, and SP-IgM began to decrease after reaching its peak at around 22-28 days, and become negative at Month 3 in 30% patients and negative at Month 7 in 79% of these patients after onset; IgG reached its peak around Day 22-28 and kept at a high level within the longest observation period for 4 months, it dropped very sharply at 7 months. The positive rates of SP-IgM and SP-IgG were higher than those of reverse transcription-polymerase chain reaction on Day 7 and 4. The established indirect ELISA has good specificity and sensitivity. IgM and IgG against SARS-CoV-2 appeared almost simultaneously in the early stage, and the level of IgG antibodies could not maintain a high plateau in the observation period of 7 months. Our data will help develop the diagnosis and vaccine of SARS-CoV-2.
Assuntos
Anticorpos Antivirais/sangue , Teste Sorológico para COVID-19/métodos , COVID-19/diagnóstico , COVID-19/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Adolescente , Adulto , Idoso , Teste de Ácido Nucleico para COVID-19/métodos , Humanos , Pessoa de Meia-Idade , Proteínas Recombinantes/imunologia , SARS-CoV-2/genética , SARS-CoV-2/isolamento & purificação , Sensibilidade e Especificidade , Glicoproteína da Espícula de Coronavírus/genética , Glicoproteína da Espícula de Coronavírus/imunologia , Fatores de Tempo , Adulto JovemRESUMO
OBJECTIVES: To characterize the MDR genomic islands (GIs) in Proteus mirabilis isolates. METHODS: Two P. mirabilis strains (C55 and C74) of chicken origin were subjected to WGS (HiSeq and PacBio) and the MDR GIs were determined. RESULTS: P. mirabilis strains C55 and C74 are clonal strains and harbour different Proteus genomic island 2 (PGI2) variants (PGI2-C55 and PGI2-C74). The MDR region of PGI2-C55 is composed of two class 1 integrons, separated by a region containing seven copies of IS26 and eight resistance genes, including blaCTX-M-3 and fosA3. The region in PGI2-C74 is a complete In4-type class 1 integron, harbouring five gene cassettes (dfrA16, blaCARB-2, aadA2, cmlA1 and aadA1). In addition, C55 and C74 carry an SXT/R391 integrative and conjugative element (ICEPmiJpn1), harbouring blaCMY-2, and a novel 50.46 kb genomic resistance island named PmGRI1-C55. PmGRI1-C55 harbours a tyrosine-type recombinase/integrase that might be responsible for the integration of PmGRI1-C55 at the 3' end of tRNA-Sec. It carries an MDR region derived from Tn2670 that harbours a Tn21 region and carries six resistance genes (catA1, blaTEM-1b, aphA1a, sul2, strA and strB). Blast analysis showed diverse PmGRI1 variants in P. mirabilis and Escherichia coli strains. CONCLUSIONS: The finding of the two new PGI2 variants highlights that the homologous recombination between shared components of class 1 integrons and transposition by IS26 promote the diversity of MDR regions in PGI2. PmGRI1 is a new GI that carries various resistance genes identified in P. mirabilis and E. coli.
Assuntos
Ilhas Genômicas , Proteus mirabilis , Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Escherichia coli , Genômica , Integrons/genética , Proteus mirabilis/genéticaRESUMO
Philadelphia chromosome (Ph)-like acute lymphoblastic leukemia (ALL) comprises â¼10% to 15% of childhood ALL cases, many of which respond exquisitely to tyrosine kinase inhibitors (TKIs), for example, imatinib in PDGFRB-rearranged ALL. However, some cases developed drug resistance to TKIs and the mechanisms are poorly understood. In this study, we identified a novel PDGFRB fusion gene, namely AGGF1-PDGFRB, and functionally characterized its oncogenic potential in vitro. Further genomic profiling of longitudinally collected samples during treatment revealed the emergence of a mutation, PDGFRBC843G , which directly conferred resistance to all generations of ABL TKIs, including imatinib, dasatinib, nilotinib, and ponatinib. PDGFRB-mutant leukemia cells are highly sensitive to multitarget kinase inhibitor CHZ868, suggesting potential therapeutic options for some patients resistant to ABL TKIs. In summary, we describe a complex clonal evolution pattern in Ph-like ALL and identified a novel PDGFRB point mutation that drives leukemia relapse after ABL TKI treatment.
Assuntos
Antineoplásicos/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Mutação , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Inibidores de Proteínas Quinases/farmacologia , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Proteínas Angiogênicas/genética , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Pré-Escolar , Humanos , Masculino , Proteínas de Fusão Oncogênica , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Inibidores de Proteínas Quinases/uso terapêutico , Recidiva , Resultado do Tratamento , Sequenciamento Completo do GenomaRESUMO
Optical clearing methods reduce the optical scattering of biological samples and thereby extend optical imaging penetration depth. However, refractive index mismatch between the immersion media of objectives and clearing reagents induces spherical aberration (SA), causing significant degradation of fluorescence intensity and spatial resolution. We present an adaptive optics method based on pupil ring segmentation to correct SA in optically cleared samples. Our method demonstrates superior SA correction over a modal-based adaptive optics method and restores the fluorescence intensity and resolution at high imaging depth. Moreover, the method can derive an SA correction map for the whole imaging volume based on three representative measurements. It facilitates SA correction during image acquisition without intermittent SA measurements. We applied this method in mouse brain tissues treated with different optical clearing methods. The results illustrate that the synaptic structures of neurons within 900 µm depth can be clearly resolved after SA correction.
Assuntos
Encéfalo/diagnóstico por imagem , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Neuroimagem/métodos , Neurônios/citologia , Óptica e Fotônica , Animais , Processamento de Imagem Assistida por Computador/métodos , Camundongos , Rede NervosaRESUMO
Observing microvasculature in its native environment provides invaluable information to understand the initiation and development of microcirculatory related diseases. However, the lack of a high-resolution three-dimensional (3D) imaging technique hinders in vivo investigation of the microvasculature. Recently, we found that the red blood cells can emit autofluorescence signals with short-wavelength two-photon excitation. In this study, we exploited this property and developed a time-resolved two-photon excitation microscopy system using a homemade 520 nm femtosecond fiber laser as the excitation source. Using this system, we could achieve intravital high-resolution 3D imaging of a microvascular network noninvasively. In a mouse tumor model, tumorous blood vessels could be observed and distinguished clearly from the normal vessels.
RESUMO
We present confocal fluorescence lifetime imaging microscopy in the second near-infrared (NIR-II) window to assess the morphological and biochemical information of live samples. A home-built superconducting single-photon detector (SSPD) was used to facilitate the NIR-II fluorescence lifetime measurement. The SSPD has many advantages, including high sensitivity to NIR-II signals (detection efficiency >50%), fast temporal response (â¼109ps), low timing jitter (â¼50ps), and low dark count rate (<100cps). We demonstrate the feasibility of the developed microscopy system by comparing fluorescence lifetimes of a range of fluorophores with emission in the NIR-II window and by performing multicolor three-dimensional fluorescence lifetime imaging of a mouse ear in vivo. The biochemical properties of the cells and tissues probed by the fluorescence lifetimes of the fluorophores provide complementary information for biomedical studies, significantly benefiting diverse applications in life science.
RESUMO
Previous experimental results show that planting spacing has significant effects on root distribution and soil suction (negative pore water pressure) due to inter-plant competition. However, there is a lack of theoretical study on this aspect. This study proposes a new physically based mathematical model to capture planting spacing effects on root growth and soil suction considering three key factors, namely hydrotropism, soil mechanical impedance and inter-plant competition. The model is mainly composed of four parts: (i) extension of root zone front; (ii) increase in root density; (iii) root water uptake and (iv) water flow in soil matrix. Root growth and root water uptake are fully coupled. In order to validate the model, laboratory and field tests were conducted on one tree (Schefflera heptaphylla) and one shrub species (Schefflera arboricola), respectively, with different planting spacings. Even though the investigated tree and shrub species had different values of leaf area index and root length density, consistent conclusions on planting spacing effects can be drawn. When planting spacing became smaller, the size of root system decreased while root density increased, hence causing higher soil suction. The model can capture the root distributions as well as induced soil suction during both evapotranspiration and rainfall events quite well for both tree and shrub species.
Assuntos
Modelos Biológicos , Raízes de Plantas , Solo , Araliaceae/crescimento & desenvolvimento , Raízes de Plantas/crescimento & desenvolvimento , Densidade Demográfica , Sucção , Árvores/crescimento & desenvolvimento , ÁguaRESUMO
This study compared the laboratory indexes in 40 non-severe COVID-19 patients with those in 57 healthy controls. In the peripheral blood system of non-severe symptom COVID-19 patients, lymphocytes, eosinophils, basophils, total procollagen type 1 amino-terminal propeptide, osteocalcin N-terminal, thyroid-stimulating hormone, growth hormone, and insulin-like growth factor-binding protein 3 significantly decreased, and total protein, albumin, alanine transaminase, alkaline phosphatase, γ-glutamyl transferase, activated partial thromboplastin time, prothrombin time, fibrinogen, D-dimer, fibrinogen degradation products, human epididymal protein 4, serum ferritin, and C-reactive protein were elevated. SARS-CoV-2 infection can affect hematopoiesis, hemostasis, coagulation, fibrinolysis, bone metabolism, thyroid, parathyroid glands, the liver, and the reproductive system.