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With the advent of hyperspectral Raman imaging technology, especially the rapid and high-resolution imaging schemes, datasets with thousands to millions of spectra are now commonplace. Standard preprocessing and regression methods such as least squares approaches are time consuming and require input from highly trained operators. Here we propose a solution to this analytic bottleneck through a convolutional neural network trained fully on synthetic data and then applied to experimental measurements, including cases where complete spectral information is missing (i.e. an underdetermined model). An advantage of the model is that it combines background correction and regression into a single step, and does not require user-selected parameters. We compare our results with traditional least squares methods, including the popular asymmetric least squares (AsLS) approach. Our results demonstrate that the proposed CNN model boasts less sensitivity to parameter selection, and with a rapid processing speed, with performance equal to or better than comparison methods. The performance is validated on synthetic spectral mixtures, as well as experimentally measured single-vesicle liposome data. [ABSTRACT FROM AUTHOR]

Thrombospondin‐1 (TSP1) is generally assumed to suppress the growth of osteosarcoma through inhibiting angiogenesis; however, it is unclear whether TSP1 could affect the antitumor immunity against osteosarcoma. We aimed to explore the immune‐related tumor‐promoting effects of TSP1 and decipher its underlying mechanism. First, we identified that TSP1 regulated programmed death‐ligand 1 (PD‐L1) expression, which was related to the CD8+ T cells anergy in osteosarcoma cells. The exact role of PD‐L1 in the immunosuppressive effect of TSP1 was then further confirmed by the addition of the PD‐L1 neutralizing Ab. With the addition of PD‐L1 neutralizing Abs during cocultivation, the inhibition of CD8+ T cells was abolished to a certain extent. Further mechanistic investigations showed that TSP1‐induced PD‐L1 upregulation was achieved by activation of the signal transducer and activator of transcription 3 (STAT3) pathway. In vivo experiments also indicated that TSP1 overexpression could promote the growth of primary lesions, whereas TSP1 knockdown effectively inhibits the growth of the primary lesion as well as lung metastasis by restoring the antitumor immunity. Thrombospondin‐1 knockdown combined with PD‐L1 neutralizing Ab achieved a more pronounced antitumor effect. Taken together, our study showed that TSP1 upregulates PD‐L1 by activating the STAT3 pathway and, therefore, impairs the antitumor immunity against osteosarcoma. [ABSTRACT FROM AUTHOR]

Raman hyperspectral imaging is a powerful method to obtain detailed chemical information about a wide variety of organic and inorganic samples noninvasively and without labels. However, due to the weak, nonresonant nature of spontaneous Raman scattering, acquiring a Raman imaging dataset is time-consuming and inefficient. In this paper we utilize a compressive imaging strategy coupled with a context-aware image prior to improve Raman imaging speed by 5- to 10-fold compared to classic point-scanning Raman imaging, while maintaining the traditional benefits of point scanning imaging, such as isotropic resolution and confocality. With faster data acquisition, large datasets can be acquired in reasonable timescales, leading to more reliable downstream analysis. On standard samples, context-aware Raman compressive imaging (CARCI) was able to reduce the number of measurements by ∼85% while maintaining high image quality (SSIM >0.85). Using CARCI, we obtained a large dataset of chemical images of fission yeast cells, showing that by collecting 5-fold more cells in a given experiment time, we were able to get more accurate chemical images, identification of rare cells, and improved biochemical modeling. For example, applying VCA to nearly 100 cells' data together, cellular organelles were resolved that were not faithfully reconstructed by a single cell's dataset. [ABSTRACT FROM AUTHOR]

The majority of problems in analytical Raman spectroscopy are mathematically over-determined, where many more spectral variables are measured than analytic outputs (such as chemical concentrations) are calculated. Thus, to improve spectral throughput and simplify system design, some researchers have explored the use of low resolution Raman systems for cell or tissue classification, achieving accuracy independent of spectral resolution. However, the tradeoffs inherent in this approach have not been systematically studied. Here, we theoretically and experimentally explore the relationship between spectral resolution and analytical error. We show that decreased spectral resolution leads to spectral signal-to-noise ratio and therefore more reliable results and lower limits of detection for equivalent integration times in blind unmixing of hyperspectral images. Our theoretical analysis demonstrates that the primary benefit of low resolution Raman spectroscopy is in overcoming detector noise (such as thermal or electronic noise). Therefore, the benefits are most pronounced when utilizing lower-grade, uncooled detectors. Therefore, using a low-cost CMOS camera we experimentally demonstrate the ability of low resolution Raman spectroscopy to achieve substantially improved imaging performance compared to fully-resolved Raman spectral imaging, paving the way for cost-effective, pervasive Raman spectroscopy. [ABSTRACT FROM AUTHOR]

In this paper we report the development of a cost-effective, modular, open source, and fully automated slide-scanning microscope, composed entirely of easily available off-the-shelf parts, and capable of bright field and fluorescence modes. The automated X-Y stage is composed of two low-cost micrometer stages coupled to stepper motors operated in open-loop mode. The microscope is composed of a low-cost CMOS sensor and low-cost board lenses placed in a 4f configuration. The system has approximately 1 micron resolution, limited by the f/# of available board lenses. The microscope is compact, measuring just 25×25×30 cm, and has an absolute positioning accuracy of ±1 μm in the X and Y directions. A Z-stage enables autofocusing and imaging over large fields of view even on non-planar samples, and custom software enables automatic determination of sample boundaries and image mosaicking. We demonstrate the utility of our device through imaging of fluorescent- and transmission-dye stained blood and fecal smears containing human and animal parasites, as well as several prepared tissue samples. These results demonstrate image quality comparable to high-end commercial microscopes at a cost of less than US$400 for a bright-field system, with an extra US$100 needed for the fluorescence module. [ABSTRACT FROM AUTHOR]

Current flow‐based blood counting devices require expensive and centralized medical infrastructure and are not appropriate for field use. In this article we report a streamlined, easy‐to‐use method to count red blood cells (RBC), white blood cells (WBC), platelets (PLT) and 3‐part WBC differential through a cost‐effective and automated image‐based blood counting system. The approach consists of using a compact, custom‐built microscope with large field‐of‐view to record bright‐field and fluorescence images of samples that are diluted with a single, stable reagent mixture and counted using automatic algorithms. Sample collection utilizes volume‐controlled capillary tubes, which are then dropped into a premixed, shelf‐stable solution to stain and dilute in a single step. Sample measurement and analysis are fully automated, requiring no input from the user. Cost of the system is minimized through the use of custom‐designed motorized components. We compare the performance of our system, as operated by trained and untrained users, to the clinical gold standard on 120 adult blood samples, demonstrating agreement within Clinical Laboratory Improvement Amendments guidelines, with no statistical difference in performance among different operator groups. The system's cost‐effectiveness, automation and performance indicate that it can be successfully translated for use in low‐resource settings where central hematology laboratories are not accessible. [ABSTRACT FROM AUTHOR]

The SOX4 transcription factor is involved in various cellular processes, such as embryonic development and differentiation. Deregulated expression of Sox4 in several human cancers has been reported to date, but its biological functions in the progression of osteosarcoma remain unclear. In this study, we found that the expression levels of SOX4 protein were significantly higher in high-grade osteosarcoma tissues and metastatic osteosarcoma tissues. Its overexpression was associated with poor prognosis in osteosarcoma. Knockdown of the SOX4 gene in the osteosarcoma cell lines resulted in decreased cell proliferation, migration, invasion, and induced apoptosis. After SOX4 gene silencing, the protein expression levels of Bax, Caspase-3, and P53 in osteosarcoma cells were significantly elevated, while the protein expression levels of Bcl-2, MMP2, and MMP9 were obviously decreased. In vivo analysis in nude mice further confirmed that knockdown of SOX4 suppressed tumor growth. In conclusion, SOX4 appears to be an important tumor oncogene in the regulation of osteosarcoma cell proliferation, apoptosis, and invasion, and it may be a potential target for effective osteosarcoma therapy. [ABSTRACT FROM AUTHOR]

Focal adhesion kinase ( FAK) overexpression is related to invasive and metastatic properties in different kinds of cancers. Target therapy by inhibiting FAK has achieved promising effect in some cancer treatments, but its effect in human osteosarcoma has not been well studied. In the present study, we analyzed the antitumor efficacy of PF562271, an FAK inhibitor, against osteosarcoma in vitro and in vivo. Phosphorylated FAK (Y397) was highly expressed in primary human osteosarcoma tumor samples and was associated with osteosarcoma prognosis and lung metastasis. PF562271 greatly suppressed proliferation and colony formation in human osteosarcoma cell lines. In addition, treatment of osteosarcoma cell lines with PF562271 induced apoptosis and downregulated the activity of the protein kinase B/mammalian target of rapamycin pathway. PF562271 also impaired the tube formation ability of endothelial cells in vitro. Finally, oral treatment with PF562271 in mice dramatically reduced tumor volume, weight, and angiogenesis of osteosarcoma xenografts in vivo. These results indicate that FAK inhibitor PF562271 can potentially be effectively used for the treatment of osteosarcoma. [ABSTRACT FROM AUTHOR]

Background: Humeral shaft fractures are generally managed with the conventional posterior open reduction and internal fixation (ORIF) or minimally invasive plate osteosynthesis (MIPO). This study was aimed at comparing the outcomes of these surgical techniques in terms of the vascular integrity of the mid-distal humeral shaft.Methods: Twelve upper limbs were harvested from 6 fresh cadavers. ORIF or MIPO was randomly performed on either side of each pair of limbs. The axillary artery was perfused with a latex-lead tetraoxide red solution to visualize the vascular structures. The vascular integrity of the humerus was examined by plain radiography and dissection. The periosteal filling achieved with each technique was scored and the scores compared.Results: In each limb, one main nutrient artery entering the mid-distal humeral shaft anteromedially (83.3 %) or medially (16.7 %) was first identified. No case of injury to the main nutrient artery was noted for either surgical technique. Injuries to the accessory nutrient arteries entering the mid-distal humeral shaft from the posterior aspect were absent in the MIPO cases, but occurred in 52.9 % of the ORIF cases. In addition, MIPO was also superior to the open plate technique showed superior periosteal filling than.Conclusions: Our results showed that the MIPO technique is superior to the ORIF in terms of preserving the vascular integrity of the mid-distal humeral shaft. [ABSTRACT FROM AUTHOR]

We propose a projective navigation system for fluorescence imaging and image display in a natural mode of visual perception. The system consists of an excitation light source, a monochromatic charge coupled device (CCD) camera, a host computer, a projector, a proximity sensor and a Complementary metal–oxide–semiconductor (CMOS) camera. With perspective transformation and calibration, our surgical navigation system is able to achieve an overall imaging speed higher than 60 frames per second, with a latency of 330 ms, a spatial sensitivity better than 0.5 mm in both vertical and horizontal directions, and a projection bias less than 1 mm. The technical feasibility of image-guided surgery is demonstrated in both agar-agar gel phantoms and an ex vivo chicken breast model embedding Indocyanine Green (ICG). The biological utility of the system is demonstrated in vivo in a classic model of ICG hepatic metabolism. Our benchtop, ex vivo and in vivo experiments demonstrate the clinical potential for intraoperative delineation of disease margin and image-guided resection surgery. [ABSTRACT FROM AUTHOR]

Purpose – The purpose of this paper is to establish a combined model for selecting key indexes of complex equipment, and then improve the cost forecasting precision of the model. The problem how to choose the key elements of complex products has always been concerned on many fields, such as cost assessment, investment decision making, etc. Design/methodology/approach – Using Grey System Theory to establish a cost estimation model of complicated equipment is more reasonable under the few data and poor information. Therefore, this paper constructs cost index’s system of complex equipment, and then quantitative and qualitative analysis methods are utilized to calculate the grey entropy between the characteristic parameter and the behavior parameters. Further, establish the grey relational clustering matrix of the behavior sequences by using the grey relative incidence analysis. Finally, the authors select key indicators according to the grey degree. Findings – The experiment demonstrates that the cost key parameters of complex equipment can be successfully screened out by the proposed approach, and the cost estimation accuracy of complicated products is improved. Practical implications – The method proposed in this paper could be utilized to solve some practical problems, particularly the selection of cost critical parameters for complex products with few samples and poor information. Taking the cost key indexes of civil aircraft as an example, the results verified the validity of the GICM model. Originality/value – In this paper, the authors develop the method of GICM model. Taking the data of civil aircraft as an example, the authors screen the key indicators of complex products successfully, and improve the prediction accuracy of the GM (1, N) model by using the selected parameters, which provides a reference for some firms. [ABSTRACT FROM AUTHOR]

Traditional line-scan Raman imaging features a rapid imaging speed while preserving complete spectral information, yet has diffraction-limited resolution. Sinusoidally structured line excitation can yield an improvement in the lateral resolution of the Raman image along the line's direction. However, given the need for the line and spectrometer slit to be aligned, the resolution in the perpendicular direction remains diffraction limited. To overcome this, we present here a galvo-modulated structured line imaging system, where a system of three galvos can arbitrarily orient the structured line on the sample plane, while keeping the beam aligned to the spectrometer slit in the detection plane. Thus, a two-fold isotropic improvement in the lateral resolution fold is possible. We demonstrate the feasibility using mixtures of microspheres as chemical and size standards. The results prove an improvement in the lateral resolution of 1.8-fold (limited by line contrast at higher frequencies), while preserving complete spectral information of the sample.

Because of the bulk, complexity, calibration requirements, and need for operator training, most current flow-based blood counting devices are not appropriate for field use. Standard imaging methods could be much more compact, inexpensive, and with minimal calibration requirements. However, due to the diffraction limit, imaging lacks the nanometer precision required to measure red blood cell volumes. To address this challenge, we utilize Mie scattering, which can measure nanometer-scale morphological information from cells, in a dark-field imaging geometry. The approach consists of a custom-built dark-field scattering microscope with symmetrically oblique illumination at a precisely defined angle to record wide-field images of diluted and sphered blood samples. Scattering intensities of each cell under three wavelengths are obtained by segmenting images via digital image processing. These scattering intensities are then used to determine size and hemoglobin information via Mie theory and machine learning. Validation on 90 clinical blood samples confirmed the ability to obtain mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and red cell distribution width (RDW) with high accuracy. Simulations based on historical data suggest that an instrument with the accuracy achieved in this study could be used for widespread anemia screening., Competing Interests: ZJS in a named inventor on US patent 10024858B2. The other authors declare no conflicts of interest., (© 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement.)

Biological nanoparticles are important targets of study, yet their small size and tendency to aggregate makes their heterogeneity difficult to profile on a truly single-particle basis. Here we present a label-free system called 'Raman-enabled nanoparticle trapping analysis' (R-NTA) that optically traps individual nanoparticles, records Raman spectra and tracks particle motion to identify chemical composition, size, and refractive index. R-NTA has the unique capacity to characterize aggregation status and absolute chemical concentration at the single-particle level. We validate the method on NIST standards and liposomes, demonstrating that R-NTA can accurately characterize size and chemical heterogeneity, including determining combined morpho-chemical properties such as the number of lamellae in individual liposomes. Applied to extracellular vesicles (EVs), we find distinct differences between EVs from cancerous and noncancerous cells, and that knockdown of the TRPP2 ion channel, which is pathologically highly expressed in laryngeal cancer cells, leads the EVs to more closely resemble EVs from normal epithelial cells. Intriguingly, the differences in EV content are found in small subpopulations of EVs, highlighting the importance of single-particle measurements. These experiments demonstrate the power of the R-NTA system to measure and characterize the morpho-chemical heterogeneity of bionanoparticles.

In this study, we identified microRNAs that regulate the expression of programmed death-ligand 1(PD-L1) in osteosarcoma and investigated their role in PD-L1-targeted immunotherapy. MicroRNA sequencing analysis showed that the expression of PD-L1 is regulated by microRNA-200a in U2OS, 143B, and K7 osteosarcoma cells. MicroRNA-200a overexpression induced the upregulation of PD-L1 in the osteosarcoma cells. CD8 + T cells co-cultured with microRNA-200a-overexpressing osteosarcoma cells showed reduced survival, proliferation, and secretion of granzyme B and perforin. The same phenomenon was also observed in the K7-derived syngeneic mouse model, as microRNA-200a promoted tumor growth by increasing the percentage of Foxp3 + regulatory T lymphocytes while reducing the proportions of CD4 + , CD8 + , and IFN-γ + cytotoxic T lymphocytes. But microRNA-200a overexpression group was also more responsive to PD-L1-targeted immunotherapy than the controls. In addition, the tumor tissues from 32 osteosarcoma patients showed that high expression of microRNA-200a and PD-L1 was associated with poor tumor necrosis rate after chemotherapy. Moreover, we confirmed that tensin homolog deleted on chromosome ten (PTEN) could act as the target gene for microRNA-200a during the upregulation of PD-L1. Thus, our findings provide important and novel insight into a regulatory axis involving microRNA-200a/PTEN/ PD-L1 axis, which determines osteosarcoma growth and the efficacy of PD-L1-targeted immunotherapy.

Biological microfibers are remarkable materials with diversity in their chemistry, structure and functions that provide a range of solutions for photonic structures. Here we proposed and demonstrated a humidity detection technique for spectral tuning of whispering gallery modes (WGMs) in a cylindrical microresonator formed by a piece of spider egg sac silk (SpEss) from Araneus Ventricosus. We launched a supercontinuum laser into the SpEss via a tapered single-mode fiber to excite WGMs. When the ambient humidity changed, the profile diameter and effective refractive index of the SpEss changed, which caused the WGM resonant dips to shift. The experimental results showed that when the relative humidity (RH) changed from 20% to 75% RH, the average testing sensitivity of the proposed sensor was 389.1 pm/%RH and the maximum testing sensitivity was 606.7 pm/%RH in the range of 60% to 75% RH. Also, the proposed SpEss-based humidity sensor showed a fast response time of 494 ms and good repeatability with fluctuations less than 8% compared with the initial test values. The SpEss-based sensor expanded the application of spider silk as a biodegradable and biocompatible material in biochemical sensing.

We propose one-step synthesis of hemoglobin-laden microcapsules (HbMs) by the liquid-driven coaxial flow focusing (LDCFF) method, in which purified hemoglobin (Hb) is fully encapsulated inside photocurable resin with a solid core-shell structure. The polymeric shell constitutes a rigid semipermeable membrane which prevents the leakage of Hb and allows oxygen transport. Gas-binding capacity and oxygen affinity of HbMs are comparable to the purified Hb. Solid HbMs phantoms (SHMPs) are also fabricated using resin-based HbMs homogeneously mixed with oil-soluble silicone. The spectral characteristics of SHMPs closely resemble to that of oxy-hemoglobin, and its long-term stability is more than 25 weeks. HbM dispersion with certain absorption and scattering properties circulating through the dialysis tube simulates tissue oxygen dynamics compared with the post occlusive reactive hyperemia (PORH) test on the healthy volunteer. The occlusion test indicates that HbMs dispersion solution can substitute conventional blood phantom. Our studies have shown that microencapsulation of Hb effectively protects it from the external environment, which makes HbMs promising for optical tissue-simulating phantom in biomedical fields.

We propose a portable phantom system for calibration and validation of medical optical devices in a clinical setting. The phantom system comprises a perfusion module and an exchangeable tissue-simulating phantom that simulates tissue oxygenation and blood perfusion. The perfusion module consists of a peristaltic pump, two liquid storage units, and two pressure suppressors. The tissue-simulating phantom is fabricated by a three-dimensional (3D) printing process with microchannels embedded to simulate blood vessels. Optical scattering and absorption properties of biologic tissue are simulated by mixing graphite powder and titanium dioxide powder with clear photoreactive resin at specific ratios. Tissue oxygen saturation (StO 2 ) and blood perfusion are simulated by circulating the mixture of blood and intralipid at different oxygenation levels and flow rates. A house-made multimodal imaging system that combines multispectral imaging and laser speckle imaging are used for non-invasive detection of phantom oxygenation and perfusion, and the measurements are compared with those of a commercial Moor device as well as numerical simulation. By acquiring multimodal imaging data from one phantom and applying the calibration factors in different settings, we demonstrate the technical feasibility to calibrate optical devices for consistent measurements. By simulating retina tissue vasculature and acquiring functional images at different tissue oxygenation and blood perfusion levels, we demonstrate the clinical potential to simulate tissue anomalies. Our experiments imply the clinical potential of a portable, low-cost, and traceable phantom standard to calibrate and validate medical optical devices for improved performance.

Raman spectroscopy is a label-free method of obtaining detailed chemical information about samples. Its compatibility with living tissue makes it an attractive choice for biomedical analysis, yet its translation from a research tool to a clinical tool has been slow, hampered by fundamental Raman scattering issues such as long integration times and limited penetration depth. In this review we detail the how combining Raman spectroscopy with other techniques yields multimodal instruments that can help to surmount the translational barriers faced by Raman alone. We review Raman combined with several optical and non-optical methods, including fluorescence, elastic scattering, OCT, phase imaging, and mass spectrometry. In each section we highlight the power of each combination along with a brief history and presentation of representative results. Finally, we conclude with a perspective detailing both benefits and challenges for multimodal Raman measurements, and give thoughts on future directions in the field., Competing Interests: The authors declare no conflict of interest.

Microenvironment at the metastatic locus usually differs greatly from that present in the site of primary tumor formation and it has a significant impact on the fate of the extravasated cancer cells. We compared gene expression signatures of primary tumors and lung metastatic tumors, and identified Thrombospondin-1 (TSP1) as highly up-regulated in the lung metastatic tumors. Immunohistochemical staining further indicated that TSP1 protein expression was higher in lung metastatic tumors compared to primary tumors in both osteosarcoma xenograft model and human clinical samples. TSP1 mRNA level is significantly associated with the Enneking stage of osteosarcoma and lung metastasis. TGF-β pathways could stimulate the TSP1 expression in osteosarcoma cells. Knockdown of TSP1 expression in osteosarcoma cells dramatically suppressed cell wound healing, migration and invasion. Treatment with recombinant TSP1 protein in osteosarcoma cells significantly promoted cell wound healing, migration and invasion. Meanwhile, suppression of TSP1 in osteosarcoma cells resulted in decreased pulmonary metastasis in vivo . Mechanistically, TSP1 increased expression of metastasis related genes, including MMP2, MMP9 and Fibronectin 1. TSP1 promoted osteosarcoma cell motility through the activation of FAK pathway. Taken together, our study provides evidence of the contributions of TSP1 to the lung metastasis of osteosarcoma and suggests that this protein may represent a potential therapeutic target for osteosarcoma lung metastasis., Competing Interests: CONFLICTS OF INTEREST We have no potential conflicts of interest to declare.

BACKGROUND Understanding the nutrient foramina is critical to clinical practice. An insult to the nutrient foramina can be caused by trauma and/or surgical dissection and lead to devascularization and bad outcomes. Few studies have looked at the humerus, and no studies have described relative information of humeral nutrient foramen related to anatomical structures that might be located by palpable landmarks. In this study, we analyzed the anatomical features of the nutrient foramina of the diaphyseal humerus and provide a discussion of clinical relevance. MATERIAL AND METHODS We dissected 19 cadavers and analyzed the relative positions of the foramina and surrounding muscles, and the number, direction, diameter, and location of the nutrient foramina. Foramina index and a new landmark index were used to calculate the location. We compared the data from both sides and the relationships between transverse and longitudinal locations, diameter and total length, and foramina index and landmark index were also analyzed. RESULTS The humeri had one or two main nutrient foramina located in a small area between the coracobrachialis and brachial muscles and oriented toward the elbow. The mean diameter was 1.11±0.32 mm. The mean index and landmark index were 43.76±4.94% and 42.26±5.35%, respectively. There were no differences between sides in terms of diameter, length, or nutrient foramina index. There were no significant correlations between transverse and longitudinal locations or diameter and total length. The foramina index and landmark index showed strong positive correlation (r=0.994, p<0.0001). CONCLUSIONS Our study provides details about the nutrient foramina that will benefit clinicians who treat injuries and diseases of the humerus. Surgeons should be mindful of soft tissue in the foraminal area during surgical procedures.