- 标题
- 摘要
- 关键词
- 实验方案
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Intraoperative Probe-Based Confocal Endomicroscopy to Histologically Differentiate Thyroid From Parathyroid Tissue Before Resection
摘要： Background. Frozen section is the standard method to histologically distinguish parathyroid tissue from thyroid tissue during endocrine neck surgery. Frozen section can be time-consuming and costly. Its drawback is that it is to be performed only after the removal of a suspected pathological tissue. This study demonstrates the use of probe-based confocal laser endomicroscopy (pCLE) to confirm histology prior to tissue resection. Design. A prospective, single-institution, nonrandomized study was conducted. No sample size calculation was performed for this observational trial. The primary objective was the description of histological rendering of normal and pathological tissues through pCLE. Real-time in vivo fluorescence microscopy imaging was performed with the CystoFlex UHD probe after intravenous injection of 2.5 mL of 10% fluorescein sodium. Results. Eleven patients with hyperparathyroidism and thyroid conditions were included. A total of 104 videos showing thyroid, parathyroid, adipose tissue, muscle, laryngeal nerve, and lymph nodes were recorded. Videos were compared with visual information and pathological samples (when sampling was indicated). Thyroid tissue could be identified based on the presence of colloid follicles (intensely fluorescent area surrounded by a small ridge of low-fluorescence epithelial cells) including the pathognomonic aspect of resorption vacuole. Parathyroid tissue could be identified based on a regular, “diamond-shaped” capillary network encompassing parathyroid chief cells. Blinded reinterpretation of pCLE videos demonstrated an 89.3% sensitivity and a 90% specificity as compared with histology in tissue recognition. Conclusion. This pilot study describes representative renderings of intraoperative pCLE to nontraumatically differentiate thyroid, parathyroid, and lymph nodes before surgical removal.
关键词： parathyroidectomy,thyroid surgery,pCLE,in vivo fluorescence microscopy,probe-based confocal laser endomicroscopy
更新于2025-09-10 09:29:36
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Classification of Limbal Stem Cell Deficiency Using Clinical and Confocal Grading
摘要： To grade the severity of limbal stem cell deficiency (LSCD) based on the extent of clinical presentation and central corneal basal epithelial cell density (BCD). This is a retrospective observational comparative study of 48 eyes of 35 patients with LSCD and 9 eyes of 7 normal subjects (controls). Confocal images of the central cornea were acquired. A clinical scoring system was created based on the extent of limbal and corneal surface involvement. LSCD was graded as mild, moderate, and severe stages based on the clinical scores. The degree of BCD reduction was given a score of 0 to 3. Compared with BCD in controls, BCD decreased by 23.0%, 40.4%, and 69.5% in the mild, moderate, and severe stages of LSCD classified by the clinical scoring system, respectively. The degree of BCD reduction was positively correlated with larger limbal and corneal surface involvement and when the central visual axis was affected (all P # 0.0005). Mean corrected distance visual acuity logarithm of the minimum angle of resolution was 0.0 6 0.0 in control eyes, 0.2 6 0.5 in mild LSCD, 0.6 6 0.4 in moderate LSCD, and 1.6 6 1.1 in severe LSCD (P , 0.0001). There was a significant correlation between a higher clinical score and corrected distance visual acuity logarithm of the minimum angle of resolution (rho = 0.82; P , 0.0001) and a greater decrease in BCD (rho = 20.78; P , 0.0001). A clinical scoring system was developed to assess the extent of clinical presentation of LSCD. A classification system to grade the severity of LSCD can be established by combining the BCD score with the clinical score.
关键词： diagnosis,classification system,limbal stem cell,in vivo confocal microscopy,limbal stem cell deficiency
更新于2025-09-09 09:28:46
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<i>In vitro</i> and <i>in vivo</i> phasor analysis of stoichiometry and pharmacokinetics using short lifetime near-infrared dyes and time-gated imaging
摘要： We introduce a simple new approach for time-resolved multiplexed analysis of complex systems using near-infrared (NIR) dyes, applicable to in vitro and in vivo studies. We show that fast and precise in vitro quantification of NIR fluorophores? short (sub-nanosecond) lifetime and stoichiometry can be done using phasor analysis, a computationally efficient and user-friendly representation of complex fluorescence intensity decays obtained with pulsed laser excitation and time-gated camera imaging. We apply this approach to the study of binding equilibria by F?rster resonant energy transfer using two different model systems: primary/secondary antibody binding in vitro and ligand/receptor binding in cell cultures. We then extend it to dynamic imaging of the pharmacokinetics of transferrin engagement with the transferrin receptor in live mice, elucidating the kinetics of differential transferrin accumulation in specific organs, straightforwardly differentiating specific from non-specific binding. Our method, implemented in a freely-available software, has the advantage of time-resolved NIR imaging, including better tissue penetration and background-free imaging, but simplifies and considerably speeds up data processing and interpretation, while remaining quantitative. These advances make this method attractive and of broad applicability for in vitro and in vivo molecular imaging, and could be extended to applications as diverse as image guided-surgery or optical tomography.
关键词： Lifetime,Fluorescence,FRET,Mouse,Phasor,in vivo,binding equilibrium,transferrin,pharmacokinetics,Near-infrared
更新于2025-09-09 09:28:46
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Photo Protection against Visible Light Induced Pigmentation
摘要： This paper presents in vivo an in vitro studies demonstrating the induction of pigmentation in human skin by visible light which can be blocked by using formulation containing the correct amount of yellow iron oxide (YIO). An in vitro absorption method was developed to determine the protection provided by a test formulation containing 4.5% YIO using an IPD UVA-VIS action spectrum. Following the development of the in vitro method and in vivo study with 10 normal healthy volunteers with Fitzpatrick skin phototypes IV to VI was conducted to verify if the predictive model. The in vitro model for visible light protection provided a protection factor of 2.5 using the in vitro absorption spectrum of 4.5% of YIO with a very similar result from the in vivo study with a protection factor of 3.0. Multiple daily exposures of visible light have shown increase in skin pigmentation and the application of YIO provide less development of pigmentation when compared to unprotected skin. In vitro testing of the absorbance of the pigmented formulation using a proposed action spectrum for immediate pigment darkening (IPD) response in the visible light range supports the in vivo protection observations for persistent pigment darkening (PPD) and can be used as predictor for skin pigmentation induced by visible light.
关键词： visible light,in vitro,in vivo,yellow iron oxide,pigmentation
更新于2025-09-09 09:28:46
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Intraoperative Thermography of the Electrical Stimulation Mapping: A Safety Control Study
摘要： A standard procedure for continuous intraoperative monitoring of the integrity of the corticospinal tracts by eliciting muscle responses is the electric stimulation mapping (ESM). However, standard ESM protocols are ineffective in 20% of young children. We have developed a novel, highly efficient paradigm consisting of short-time burst (30 ms) of high frequency (500 Hz) and high peak current (≤100 mA), which may cause local tissue overheating. The presented safety control study was therefore designed. The infrared thermography camera captured to-be-resected cortex of 13 patients in vivo during ESM. Thermograms were image processed to reveal discrete ESM thermal effect of currents from 10 to 100 mA. Peak 100 mA currents induced a maximal increase in temperature of 3.1 °C, 1.23 ± 0.72 °C in average. The warming correlated with stimulating electrode resistance (p < 0.001). The measurement uncertainty was estimated ±1.01 °C for the most skeptical conditions. The histopathological evaluation of stimulated tissue (performed in all cases) did not show any destructive changes. Our study demonstrates the ability of the thermographic camera to measure the discrete thermal effect of the ESM. The results provide evidence for the safety of the proposed protocol for full range currents with minimal risk of brain tissue damage.
关键词： neurosurgery,safety control,Electrical stimulation mapping,thermography,thermal effect,in vivo
更新于2025-09-09 09:28:46
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Molecular Imaging: <i>In Vivo</i> Agents for the Diagnosis and Treatment of Cancer
摘要： Molecular imaging continues to advance the goal of improving diagnosis and treatment in cancer. This special issue examines a cross section of the current basic and clinical research across imaging modalities, probes, and molecular targets. The issue articles permit comparison of the advantages and limitations of varied modalities, including PET, SPECT, CT, MRI, ultrasound imaging, and fluorescence imaging. The corresponding agents in development are able to interact with many targets of relevance to cancer. Highly sensitive and specific in vivo agents permit visualization of receptor systems, enzymes, and proteins involved in cancer initiation, maintenance, and spread. As another dimension of cancer targeting, molecular probes for the tumor microenvironment, including stromal, endothelial, and immune cells, are increasingly recognized as key factors in attacking cancer. The caleidoscope of targets and methods available to achieve these goals is exemplified in this special issue. The topics cover the development of molecular imaging agents targeting the cholecystokinin 2 receptor (CCK2R), adenosine A3 receptor (A3R), and the human epidermal growth factor receptor 2 (HER2). Cancer stroma targeting is addressed with molecular probes for the vascular cell adhesion molecule 1 (VCAM-1) and endoglin (CD105), a proangiogenic growth factor, which are both overexpressed in a variety of malignancies. The tools used in the selection of displayed articles were intact molecules, small peptides, engineered proteins, antibodies and antibody fragments, nanoparticles, and targeting microbubbles, demonstrating the breadth of scaffolds currently to investigators and soon to cancer clinicians. Each paper included in this special issue approaches the challenge of molecular imaging in diagnosis and treatment of cancer in a different way, focusing on new molecules, innovative labelling strategies, and characterization in disease models. Each contribution stands on its own as a marker for the next advances in cancer understanding and patient care. The editors have endeavored to have the authors clearly point up the advantages of the current technologies and demonstrate the needs for advancement to the next stages of development. We hope this effort will stimulate the reader to think hard about their own research and how the community of cancer imaging investigators can continue progress from proof of concept to clinical use.
关键词： Cancer,Treatment,Diagnosis,In Vivo Agents,Molecular Imaging
更新于2025-09-09 09:28:46
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Comparative Study of Tissue Distribution of Chlorin e6 Complexes with Amphiphilic Polymers in Mice with Cervical Carcinoma
摘要： Many photosensitizers, including chlorins, are highly hydrophobic, which makes intravenous administration problematic and affects their delivery to the tumor and uptake in the cells. Moreover, self-aggregation of the photosensitizer in aqueous solution reduces fluorescence quantum yield, triplet state, and singlet oxygen generation, and consequently diminishes photosensitizing activity. To address these issues, it was proposed to use biocompatible water-soluble polymers. However, animal studies of the photosensitizer-polymer systems are still very limited. In this work, polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), and pluronic F108 were used for dissolution of chlorin e6 (Ce6). Dynamics of accumulation of the formulations in a mouse cervical carcinoma and clearance from normal tissue, drug plasma concentrations and tissue distribution after intravenous injection were investigated. Ce6 alone and clinically used photosensitizer Photoditazine served as a control. The results showed that none of the polymers significantly changed fluorescence kinetics in the tumor. Concentration of the Ce6 formulated with polymers in the tumor tissue was comparable with Photoditazine, but uptake in the skin was less. At the same time, tumor-to-skin ratios of the Ce6-polymer complexes were similar to free Ce6. We concluded that the use of the polymeric formulation is reasonable for fluorescence diagnosis and PDT of cancer.
关键词： Chemical extraction,Amphiphilic polymer,Mouse cervical carcinoma,Photosensitizer,Chlorin e6,PDT,Fluorescence imaging in vivo
更新于2025-09-09 09:28:46
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Colorimetric and NIR fluorescence probe with multiple binding sites for distinguishing detection of Cys/Hcy and GSH in vivo
摘要： Although some progress has been made in distinguishing detection of biothiols, NIR biothiol fluorescent probes for simultaneous distinguishing detection of Cys, Hcy and GSH in vivo have not been reported. The design of these probes involves the introduction of NIR fluorophores and multiple binding sites, so the integrated design of probes remains a challenge. Although Cys, Hcy and GSH have the common functional group: a sulfydryl group and an amino group, due to their differences in spatial structure, they may react with multiple binding sites probes to produce different reaction products in different bonding mechanisms, resulting in the different colors and fluorescent signals changes of the system. Therefore, multiple binding sites fluorescent probes can realize their discrimination detection. For NIR fluorescent probe, it is easier to realize in vivo imaging to promote the research of biothiols in clinical diagnosis. In our work, not only multiple binding sites were constructed in the compound, but also NIR fluorophores were introduced. This enables the probe to not only efficiently distinguish detection of Cys/Hcy and GSH but also achieve fluorescence imaging in vivo. We believe this result is a milestone in the discrimination detection of biothiols.
关键词： Cys/Hcy,in vivo,NIR fluorescence probe,GSH,Colorimetric,multiple binding sites
更新于2025-09-09 09:28:46
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Diagnostic accuracy of ex vivo fluorescence confocal microscopy for Mohs surgery of basal cell carcinomas: a prospective study on 753 margins
摘要： Background: Frozen histological sections are currently used for intra-operative margin assessment during Mohs surgery. Fluorescence confocal microscopy (FCM) is a new tool, which offers a promising and faster alternative to frozen histology. Objective: The aim of the present study is to prospectively evaluate in a clinical setting, the accuracy of FCM as compared to frozen sections in BCC’s margin assessment. Methods: Patients with BCC, scheduled for Mohs surgery were prospectively enrolled. Freshly-excised surgical specimens were first examined through FCM and then frozen sections were evaluated. Permanent sections were finally obtained, in order to validate our sample technique. A blind re-evaluation was also performed for discordant cases between FCM and frozen sections. Sensitivity and specificity levels, as well as positive and negative predicting values were calculated and ROC curves were generated. Results: We enrolled 127 BCCs in as many patients (40.2% females). A total number of 753 sections were examined. All BCCs were located on the head and neck area. When evaluating the performance of FCM as compared to frozen sections 79.8% sensitivity, 95.8% specificity, 80.5% positive predicting and 95.7% negative predicting values were found (Area Under the Curve: .88, 95%CI .84-.92; P<.001). A total of 49 discordant cases between FCM and frozen sections evaluations were blindly re-evaluated, of which 24 were false positive and 25 false negative. The performance of FCM and frozen sections was also evaluated according to the final histopathological assessment. Conclusions: We found high levels of accuracy for FCM as compared to frozen sections evaluation, in intra-operative BCC’s margin assessment during Mohs surgery. Some technical issues still prevent a wide use of this technique, but new upcoming devices promise to overcome these limitations.
关键词： basal cell carcinoma,fluorescence confocal microscopy,Mohs surgery,skin cancer,ex vivo confocal microscopy
更新于2025-09-09 09:28:46
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<i>In vivo</i> monitoring of intracellular Ca <sup>2+</sup> dynamics in the pancreatic β-cells of zebrafish embryos
摘要： Assessing the response of pancreatic islet cells to glucose stimulation is important for understanding β-cell function. Zebrafish are a promising model for studies of metabolism in general, including stimulus-secretion coupling in the pancreas. We used transgenic zebrafish embryos expressing a genetically-encoded Ca2+ sensor in pancreatic β-cells to monitor a key step in glucose induced insulin secretion; the elevations of intracellular [Ca2+]i. In vivo and ex vivo analyses of [Ca2+]i demonstrate that β-cell responsiveness to glucose is well established in late embryogenesis and that embryonic β-cells also respond to free fatty acid and amino acid challenges. In vivo imaging of whole embryos further shows that indirect glucose administration, for example by yolk injection, results in a slow and asynchronous induction of β-cell [Ca2+]i responses, while intravenous glucose injections cause immediate and islet-wide synchronized [Ca2+]i fluctuations. Finally, we demonstrate that embryos with disrupted mutation of the CaV1.2 channel gene cacna1c are hyperglycemic and that this phenotype is associated with glucose-independent [Ca2+]i fluctuation in β-cells. The data reveal a novel central role of cacna1c in β-cell specific stimulus-secretion coupling in zebrafish and demonstrate that the novel approach we propose – to monitor the [Ca2+]i dynamics in embryonic β-cells in vivo – will help to expand the understanding of β-cell physiological functions in healthy and diseased states.
关键词： GCaMP6s,glucose-sensing of beta cells,early zebrafish development,cacna1c,in vivo imaging,Cav1.2 channel
更新于2025-09-09 09:28:46