Distribution Of Microvessels Research Articles

Mathematical modeling of 18F-Fluoromisonidazole (18F-FMISO) radiopharmaceutical transport in vascularized solid tumors

18F-Fluoromisonidazole (18F-FMISO) is a highly promising positron emission tomography radiopharmaceutical for identifying hypoxic regions in solid tumors. This research employs spatiotemporal multi-scale mathematical modeling to explore how different levels of angiogenesis influence the transport of radiopharmaceuticals within tumors. In this study, two tumor geometries with heterogeneous and uniform distributions of capillary networks were employed to incorporate varying degrees of microvascular density. The synthetic image of the heterogeneous and vascularized tumor was generated by simulating the angiogenesis process. The proposed multi-scale spatiotemporal model accounts for intricate physiological and biochemical factors within the tumor microenvironment, such as the transvascular transport of the radiopharmaceutical agent, its movement into the interstitial space by diffusion and convection mechanisms, and ultimately its uptake by tumor cells. Results showed that both quantitative and semi-quantitative metrics of 18F-FMISO uptake differ spatially and temporally at different stages during tumor growth. The presence of a high microvascular density in uniformly vascularized tumor increases cellular uptake, as it allows for more efficient release and rapid distribution of radiopharmaceutical molecules. This results in enhanced uptake compared to the heterogeneous vascularized tumor. In both heterogeneous and uniform distribution of microvessels in tumors, the diffusion transport mechanism has a more pronounced than convection. The findings of this study shed light on the transport phenomena behind 18F-FMISO radiopharmaceutical distribution and its delivery in the tumor microenvironment, aiding oncologists in their routine decision-making processes.

The Role of Microvascular Variations in the Process of Intervertebral Disk Degeneration and Its Regulatory Mechanisms: A Literature Review.

Microvascular changes are considered key factors in the process of intervertebral disk degeneration (IDD). Microvascular invasion and growth into the nucleus pulposus (NP) and cartilaginous endplates are unfavorable factors that trigger IDD. In contrast, the rich distribution of microvessels in the bony endplates and outer layers of the annulus fibrosus is an important safeguard for the nutrient supply and metabolism of the intervertebral disk (IVD). In particular, the adequate supply of microvessels in the bony endplates is the main source of the nutritional supply for the entire IVD. Microvessels can affect the progression of IDD through a variety of pathways. Many studies have explored the effects of microvessel alterations in the NP, annulus fibrosus, cartilaginous endplates, and bony endplates on the local microenvironment through inflammation, apoptosis, and senescence. Studies also elucidated the important roles of microvessel alterations in the process of IDD, as well as conducted in-depth explorations of cytokines and biologics that can inhibit or promote the ingrowth of microvessels. Therefore, the present manuscript reviews the published literature on the effects of microvascular changes on IVD to summarize the roles of microvessels in IVD and elaborate on the mechanisms of action that promote or inhibit de novo microvessel formation in IVD.

Arterial supply and morphological characteristics of sympathetic neurons in the human superior cervical ganglion.

The aim of this study was the micromorphological analysis of the distribution of microvessels, mast cells and ganglionic neurons in two parts, proximal and distal of the human superior cervical sympathetic ganglions (SCSGs). Statistical analyses were applied to detect the possible metric regional differences in their densities. Five injected human SCSGs with colored India ink and gelatin were microdissected and examined. Second group of five human SCSGs was prepared and serially sliced for CD34 and mast cell tryptase immunostaining. The microscopic fields of two parts of the SCSGs were analyzed for the following quantifications: microvessel density (MVD), mast cell density (MCD), and ganglionic cell count and measurements. The mean number of CD34-positive microvessels in microscopic fields, the MVD, had a value of 83 for the upper parts, and 82.7 for the lower parts of SCSGs. The mean number of tryptase-positive mast cells in microscopic fields, the MCD, was 4.5 in the proximal parts, and 4.7 in the distal parts of SCSGs. The mean number of ganglionic neurons in microscopic fields was 19.5 in the proximal parts, and 19.8 in the distal parts of SCSGs. The density of CD34-positive microvessels, the density of tryptase-positive mast cells, and the density, mean diameters and mean areas of ganglionic neurons were not significantly different in two observed parts, upper and lower of the SCSGs. In conclusion, the distributions of microvessels, mast cells, and neurons in two parts of the SCSGs were uniform with no specific micromorphological variations, there is a homogenous vascular and cellular pattern within the SCSGs.

Prediction of lymph node status in stage T1 esophageal squamous cell carcinoma.

263 Background: With the development of endoscopic therapy and the introduction of immunotherapies, it is more important for preserving the esophagus and uninvaded lymph nodes to avoid the damages involving the long-term quality of life and normal immune function caused by esophagectomy with radical lymphadenectomy. Methods: We collected patients with T1 ESCC who underwent endoscopic resection or esophagectomy from 2014 to 2021 in three cancer centers. The primary end point was presence of lymph node metastasis (LNM) in surgical resection specimens or detection of metastases during follow-up. Univariate and multivariate logistic regression were used to screen the clinicopathological features related to LNM. Using the function 'lrm' constructed the prediction model. The area under the curve (AUC) of receiver operating characteristic and calibration graphs were used to assess the performance of the model. An external validation was conducted to evaluate the generalization of the model. We then prospectively collected 28 surgically resected tissue samples of T1 ESCC to further supplement the model from the cellular and molecular level, and used 6 markers (CK, CD34, D2-40, CD4, CD8, FOXP3) and DAPI dye to process multiple immunofluorescence (mIF) staining. Afterwards, we analyzed the density, spatial distribution of microvessels, lymphatic capillaries and tumor-associated immune cells and the interaction between them and malignant cells in the tumor microenvironment of T1 ESCC to explore the relationship between tumor microenvironment and risk of LNM. Results: 1109 patients with T1 ESCC were included. 653 patients from two centers constituted the training cohort, and 456 patients from the third center served as the validation cohort. Multivariate logistic regression based on the training cohort showed that tumor invasion into the submucosa, poor histological grade, and lymphovascular invasion were risk factors for LNM in T1 ESCC. Subsequently, we constructed a model based on three variables to predict the risk of LNM in T1 ESCC. The model demonstrated good discriminative ability (AUC 0.858, 95%CI 0.826-0.890). After mIF staining of 28 prospectively collected T1 ESCC tissue samples, we observed that microvessels and lymphatic capillaries were abundant in the mucosa in lymph node-positive samples but not in lymph node-negative samples. In addition, the average density of FOXP3+ T cells in samples with and without metastasis was 253/mm2 and 26/mm2 (P < 0.001), and the average density of CD8+ T cells was 112/mm2 and 432/mm2 (P < 0.001) in quantitative analysis, respectively. Conclusions: This study (RENMIN-237) developed a well-performing LNM risk stratification model of T1 ESCC to reduce both overtreatment and undertreatment, and we found that FOXP3+ T cells in the tumor microenvironment are a risk factor for LNM, while CD8+ T cells can serve as a protective factor against LNM.

A Dual-Branch Cross-Modality-Attention Network for Thyroid Nodule Diagnosis Based on Ultrasound Images and Contrast-Enhanced Ultrasound Videos.

Contrast-enhanced ultrasound (CEUS) has been extensively employed as an imaging modality in thyroid nodule diagnosis due to its capacity to visualise the distribution and circulation of micro-vessels in organs and lesions in a non-invasive manner. However, current CEUS-based thyroid nodule diagnosis methods suffered from: 1) the blurred spatial boundaries between nodules and other anatomies in CEUS videos, and 2) the insufficient representations of the local structural information of nodule tissues by the features extracted only from CEUS videos. In this paper, we propose a novel dual-branch network with a cross-modality-attention mechanism for thyroid nodule diagnosis by integrating the information from tow related modalities, i.e., CEUS videos and ultrasound image. The mechanism has two parts: US-attention-from-CEUS transformer (UAC-T) and CEUS-attention-from-US transformer (CAU-T). As such, this network imitates the manner of human radiologists by decomposing the diagnosis into two correlated tasks: 1) the spatio-temporal features extracted from CEUS are hierarchically embedded into the spatial features extracted from US with UAC-T for the nodule segmentation; 2) the US spatial features are used to guide the extraction of the CEUS spatio-temporal features with CAU-T for the nodule classification. The two tasks are intertwined in the dual-branch end-to-end network and optimized with the multi-task learning (MTL) strategy. The proposed method is evaluated on our collected thyroid US-CEUS dataset. Experimental results show that our method achieves the classification accuracy of 86.92%, specificity of 66.41%, and sensitivity of 97.01%, outperforming the state-of-the-art methods. As a general contribution in the field of multi-modality diagnosis of diseases, the proposed method has provided an effective way to combine static information with its related dynamic information, improving the quality of deep learning based diagnosis with an additional benefit of explainability.

Neutrophil extracellular traps promote angiogenesis in gastric cancer

Although antiangiogenic therapy has been used in gastric cancer, disease progression due to drug resistance remains common. Neutrophils play an important role in the occurrence and progression of cancer via neutrophil extracellular traps (NETs). However, few studies have investigated angiogenic regulation in gastric cancer. We aimed to determine the role of NETs in promoting angiogenesis in gastric cancer. Multiple immunohistochemical staining was used to analyze the spatial distribution of NETs and microvessels in patient tissue samples. A mouse subcutaneous tumor model was established to determine the effect of NETs on tumor growth, and changes in microvessel density were observed via immunohistochemical staining. We screened differentially expressed proteins in HUVECs stimulated by NETs via proteomics. Cell Counting Kit-8, EdU labeling, and tubule formation assays were used to verify the effect of NETs on HUVEC proliferation, migration, and tubule formation. Blocking NETs, which was related to decreased microvessel density, significantly inhibited tumor growth in the murine subcutaneous tumor model. Compared with those of the control group, tumor volume and mass among mice in the inhibition group decreased by 61.3% and 77.9%, respectively. The NET-DNA receptor CCDC25 was expressed in HUVECs, providing a platform for NETs to promote HUVEC proliferation, migration, and tubulation. In an in vitro rat aortic explant model, NETs induced HUVEC proliferation, survival, and chemotaxis, which were not significantly different from those observed in the VEGF stimulation group. Our results confirm that NETs promote angiogenesis in gastric cancer, providing a theoretical basis for identifying new anti-vascular therapeutic targets.Graphical 1Sf8jYXSegxbAWaiHcqZYYVideo

Extracellular matrix and vascular dynamics in the kidney of a murine model for Marfan syndrome.

Fibrillin-1 is a pivotal structural component of the kidney's glomerulus and peritubular tissue. Mutations in the fibrillin-1 gene result in Marfan syndrome (MFS), an autosomal dominant disease of the connective tissue. Although the kidney is not considered a classically affected organ in MFS, several case reports describe glomerular disease in patients. Therefore, this study aimed to characterize the kidney in the mgΔlpn-mouse model of MFS. Affected animals presented a significant reduction of glomerulus, glomerulus-capillary, and urinary space, and a significant reduction of fibrillin-1 and fibronectin in the glomerulus. Transmission electron microscopy and 3D-ultrastructure analysis revealed decreased amounts of microfibrils which also appeared fragmented in the MFS mice. Increased collagen fibers types I and III, MMP-9, and α-actin were also observed in affected animals, suggesting a tissue-remodeling process in the kidney. Video microscopy analysis showed an increase of microvessel distribution coupled with reduction of blood-flow velocity, while ultrasound flow analysis revealed significantly lower blood flow in the kidney artery and vein of the MFS mice. The structural and hemodynamic changes of the kidney indicate the presence of kidney remodeling and vascular resistance in this MFS model. Both processes are associated with hypertension which is expected to worsen the cardiovascular phenotype in MFS.

INOCA affects more than the coronaries

Abstract Background Ischaemia with normal coronary arteries (INOCA) may result in disabling symptoms and has an association with adverse long-term prognosis. The diagnosis of INOCA necessitates invasive coronary angiography to perform a physiological evaluation of microvascular function. The conjunctiva has a readily assessable microvascular network in which physiological parameters can be evaluated. We compared conjunctival haemodynamics in patients with and without coronary microvascular disease (MVD) to assess if systemic microvascular dysfunction was present in this coronary artery disease sub-group. Methods In this study, we recruited patients undergoing invasive coronary angiography for the investigation of angina or angina equivalent symptoms. All patients had physiologically insignificant epicardial disease (FFR≥0.80) and underwent a physiological evaluation of coronary microvascular function. We compared a group with evidence of coronary MVD (IMR≥25 or CFR<2.0); to a group of controls without MVD (IMR<25 and CFR≥2.0). The conjunctival microvasculature was imaged using a previously validated combination of a smartphone and slit-lamp biomicroscope. The conjunctival vasculature was assessed using a semi-automated process of vessel diameter measurement and erythrocyte tracking to obtain haemodynamic parameters of microvascular function. Results A total of 111 patients were included (43 MVD and 68 controls). There were no differences in baseline demographics, co-morbidities, epicardial coronary disease severity or regular pharmacological therapies between the groups. Mean coronary flow reserve (CFR) was lower and mean index of microcirculatory resistance (IMR) higher in the MVD cohort (CFR 2.5±1.3 vs 5.2±2.5, p<0.001 and IMR 28.4±11.8 vs 13.7±5.0, p<0.001). A total of 2295 conjunctival vessels were analysed. The mean number of vessels per patient was 21.0±12.8 (3.2±3.5 arterioles and 14.8±10.8 venules). Significant reductions in axial/cross-sectional velocity, wall shear rate and wall shear stress were observed in the MVD cohort. Table 1 demonstrates a comparison of conjunctival physiological parameters between the groups. The most marked differences were observed in conjunctival arterioles. Due to the heterogenous size distribution of microvessels, arterioles were categorised into 2 diameter sub-groups (10–25 μm and 25–40 μm) for analysis (Table 2). Conclusion The reductions in microvascular blood flow velocity and rate that form the basis for the diagnosis of coronary microvascular dysfunction can be observed non-invasively in the bulbar conjunctiva microcirculation. Conjunctival vascular imaging may have utility as a non-invasive imaging modality to both diagnose microvascular dysfunction and augment conventional cardiovascular risk stratification. Funding Acknowledgement Type of funding sources: Foundation. Main funding source(s): Belfast Trust Heart Trust Fund and Northern Ireland Chest Heart and Stroke

The heterogeneous subclones might be induced by cycling hypoxia which was aggravated along with the luminal A tumor growth

BackgroundIn process of infiltrating growth in solid tumor, the tumor cells undergo a periodic hypoxia/reoxygenation (circulating hypoxia) microenvironment, which might be one of principle reasons to promote formation of tumor heterogeneity and treatment failure. Therefore, it is very important to study how the microenvironment of circulating hypoxia induces the heterogeneous subclones in tumors. Material and MethodsThe maximum cross-section of luminal A tumor (LAT) was selected for the expression of hypoxia inducible factor-1 alpha (HIF-1 alpha) and estrogen receptor-alpha (ER-alpha), and the correlation between it and tumor diameter was observed. The distribution of intratumoral micro-vessels were analyzed by 3D reconstruction and CD34 staining. A circulating hypoxia model of MCF-7 was established to detect the HIF-1alpha and ER-alpha. ResultsThere was a negative correlation between the expressions of ER-alpha and HIF-1alpha (c=−2.40; p = 0.044) in the LAT. As shown by 3D ultrasound image, there were less functional micro-vessels in the center than the periphery of tumor(P < 0.05). ER-alpha expression gradually decreased with the time course of cycling hypoxia, which is inversely related to the expression of HIF-1alpha. ConclusionLAT is composed of heterogeneous subclone cells which can be distinguished by ER-alpha and HIF-1alpha, which was closely related with cycling hypoxia microenvironment.

Abstract 9386: Marfan Syndrome Mouse Model MgΔ Lpn Presents Alterations in Glomerular Architecture and Increased Renal Vascular Resistance. Losartan Treatment Attenuates These Phenotypes

Introduction: Previously, we have shown that mgΔ lpn murine Marfan syndrome (MFS) model with a fibrillin-1 disruption presents increased Metalloproteinase-9 (MMP-9) expression in the cilliary body in eye. MMP-9 has been related to microfibrils degradation. In this study, we examined the fibrillin-1 and MMP-9 distribution in the kidney and their influence on hemodynamics. Hypothesis: The alteration of fibrillin-1 impacts the glomerular architecture and modify renal hemodynamics. Methods: Female mice at 4 months of age were used: MFS mgΔ lpn mice (n=5); mgΔ lpn treated with losartan for 3 months (n=5); and control wild type mice (n=5). The glomerular structure of the kidney was examined by histology. The distribution and quantification of fibrillin-1 and MMP-9 were analyzed by immunofluorescence assay. Blood flow in the aorta and renal artery was monitored with the probe 2SB-T206 of the Transonic Systems, and the cortical micro-hemodynamic of the kidney was examined by SDF video-microscopy (Sidestream Dark Field Imaging). Results: The mgΔ lpn mice showed reduction of fibrillin-1 deposition and an increase in MMP-9 in the entire kidney tissue, including the glomerular structures. In histologic aspects, the glomerulus showed a reduction of the glomerular area, urinary space, and vascular pole area. The reduced glomerulus dimension possibly squeezes the arteriolar system with a subsequent increase of their vascular resistance. In addition, the cortical microvessels showed a significant reduction in blood flow, and there was an increase in the distribution of microvessels with diameters 6 to 8 μm. Blood flow analysis in the mgΔ lpn showed decreased artery renal flow but did not reveal a difference in aorta blood flow when compared to WT animals. These results indicate vascular resistance in the renal artery. Losartan therapy increased fibrillin-1 and decreased MMP-9 quantifications, improved the glomerulus structures and significantly increased the velocity of the microcirculation blood flow. Conclusions: Alterations in fibrillin-1 modify the glomerular structures and impair kidney hemodynamics. Losartan therapy minimized the renal phenotypes.

Functional Surgery for Subungual Melanoma: Surgical Tips Based on Histological Analysis of 21 Cadavers.

Amputation has been the standard treatment for subungual melanoma. Although there is growing attention toward a more conservative functional surgery, specific operative techniques are not yet available. We aimed to provide objective measurements for use in functional surgery by analyzing the anatomy of the nail apparatus obtained from 21 cadavers. Nailbed thickness was histologically measured in each subunit, and skin surface anatomy was evaluated to determine the proximal resection margin. Immunohistochemical staining was performed to analyze microvessel distribution according to the nail subunit. The nailbed thickness was the thinnest at the most proximal point of the nail matrix (thumbs, 1.10 ± 0.42 mm; big toes, 1.15 ± 0.37 mm) and the thickest at the hyponychium (thumbs, 2.86 ± 0.82 mm; big toes, 2.72 ± 0.84 mm). The distance from the eponychium to the skin surface closest to the bony cortex of extensor tendon insertion was 6.92 ± 5.13 mm in thumbs and 5.14 ± 1.59 mm in big toes. The median microvessel density was the highest at the hyponychium (25.74 vessels/mm2) and lowest at the germinal matrix (16.26 vessels/mm2) (p < .05). This histological study offers practical tips, including those to help decide the proximal and deep resection margins, in functional surgery.

A prospective study of the effect and mechanism of autologous platelet-rich plasma combined with Meek microskin grafts in repairing the wounds of limbs in severely burned patients

Objective: To observe the effect of autologous platelet-rich plasma (PRP) combined with Meek microskin grafts in repairing the wounds of limbs in severely burned patients, and to explore the mechanism. Methods: The prospective controlled research method was used. From September 2016 to January 2020, 16 patients aged 18-69 years, with extensive deep burns, including 9 males and 7 females, who met the selection criteria were admitted to the Department of Burns and Plastic Surgery of the 909th Hospital of the Joint Logistic Support Force of PLA. The bilateral limbs with similar injury in 8 patients were divided into Meek skin grafting+PRP group and Meek skin grafting alone group according to the random number table; in the other 8 patients, the limbs with severer injury were included in Meek skin grafting+PRP group, and the limbs on the other side were included in Meek skin grafting alone group. The wounds of affected limbs in the two groups were treated correspondingly. On post surgery day (PSD) 10, the survival and fusion of Meek microskin grafts were observed and the survival rate and fusion rate were calculated; the histological morphology and the angiogenesis of the basal tissue of Meek microskin graft were observed by hematoxylin-eosin staining and immunohistochemical staining, respectively, with the microvessels being counted. Data were statistically analyzed with paired sample t test. Results: On PSD 10, the wounds of affected limbs in Meek skin grafting+PRP group were dry, and most of the transplanted skin grafts were closely adhered to the basal tissue; while a small amount of exudate could be found in the wounds of affected limbs in Meek skin grafting alone group, and a small part of the transplanted microskin grafts fell off or poorly attached to the basal tissue. On PSD 10, the survival rate and the fusion rate of Meek microskin grafts in the wounds of affected limbs in Meek skin grafting+PRP group were (94±3)% and (86±4)%, which were significantly higher than (89±4)% and (79±4)% of Meek skin grafting alone group, respectively (t=3.633, 4.229, P<0.01). On PSD 10, the basal epidermis was closely connected with dermis of Meek microskin grafts in the wounds of affected limbs in Meek skin grafting+PRP group, with more inflammatory cell infiltration and active microvascular hyperplasia, while the basal epidermis was less closely connected with dermis of Meek microskin grafts in the wounds of affected limbs in Meek skin grafting alone group, with obvious degeneration of collagen fibers under the dermis, less inflammatory cell infiltration, and slightly poor microvascular hyperplasia. On PSD 10, the distribution of microvessels in basal tissue of Meek microskin grafts in the wounds of affected limbs in Meek skin grafting+PRP group were densely clustered, while the distribution of microvessels in Meek skin grafting alone group were scattered, sparse, and dotted. On PSD 10, the number of microvessels in basal tissue of Meek microskin grafts in the wounds of affected limbs in Meek skin grafting+PRP group was 36±6 in each 400-fold visual field, which was significantly more than 29±7 of Meek skin grafting alone group (t=2.671, P<0.05). Conclusions: Autologous PRP can effectively promote the survival rate and fusion rate of Meek microskin grafts in the wounds of limbs after escharectomy in severely burned patients by promoting angiogenesis at the base of Meek microskin grafts.

Identify the triple-negative and non-triple-negative breast cancer by using texture features of medicale ultrasonic image: A STROBE-compliant study.

The study aimed to explore the value of ultrasound (US) texture analysis in the differential diagnosis of triple-negative breast cancer (TNBC) and non-TNBC.Retrospective analysis was done on 93 patients with breast cancer (35 patients with TNBC and 38 patients with non-TNBC) who were admitted to Taizhou people's hospital from July 2015 to June 2019. All lesions were pathologically proven at surgery. US images of all patients were collected. Texture analysis of US images was performed using MaZda software package. The differences between textural features in TNBC and non-TNBC were assessed. Receiver operating characteristic curve analysis was used to compare the diagnostic performance of textural parameters showing significant difference.Five optimal texture feature parameters were extracted from gray level run-length matrix, including gray level non-uniformity (GLNU) in horizontal direction, vertical gray level non-uniformity, GLNU in the 45 degree direction, run length non-uniformity in 135 degree direction, GLNU in the 135 degree direction. All these texture parameters were statistically higher in TNBC than in non-TNBC (P <.05). Receiver operating characteristic curve analysis indicated that at a threshold of 268.9068, GLNU in horizontal direction exhibited best diagnostic performance for differentiating TNBC from non-TNBC. Logistic regression model established based on all these parameters showed a sensitivity of 69.3%, specificity of 91.4% and area under the curve of 0.834.US texture features were significantly different between TNBC and non-TNBC, US texture analysis can be used for preliminary differentiation of TNBC from non-TNBC.

Immunohistochemical analysis of the arterial supply and mast cells of the trigeminal ganglion

The aim of this study was to quantify the distribution of microvessels and mast cells in all three parts of the trigeminal ganglion (TG). Statistical analyses were applied to investigate possible micromorphological regional differences in their density. Five serially sectioned human TGs were prepared for CD34 and mast cell tryptase immunostaining. The following quantifications were performed in microscopic fields of three parts of the TG: microvessel density (MVD), mast cell density (MCD) and ganglionic cell count. The density of CD34-positive microvessels was not significantly different in any of the three observed parts of the TG. The distribution of neurons showed no significant statistical difference in three parts of the TG. There was no difference in the density of tryptase-positive mast cells within the TG, but there was an abundant presence of mast cells in the periganglionic dural and subdural tissues, a finding hitherto not reported. We can say that there is a homogenous vascular pattern within the TG which excludes local predominance in pathogenesis of trigeminal neuralgia. Second, and more important, the finding of peri-trigeminal mast cells indicates their important role in migraine pain and confirms their degranulation as the main therapeutic goal for this condition.

Brain micro‐vasculature imaging: An unsupervised deep learning algorithm for segmenting mouse brain volume probed by high‐resolution phase‐contrast X‐ray tomography

AbstractHigh‐throughput synchrotron‐based tomographic microscopy at third generation light sources allows to probe cm‐sized samples at micrometer‐resolution. In this work, we present an approach to image a full mouse brain. With Indian‐ink as a contrast agent, it was possible to obtain 3D distribution of microvessels while a computational framework automatically extracted the morphological and geometrical embedding of the putative vascular systems. Results demonstrate the potentiality of the proposed methodology to visualize and quantify in 3D details of the brain tissue with an image quality and resolution previously unachievable.

Correlation Between Proteolytic Enzymes and Microangiogenesis in Degenerative Intervertebral Disc Nucleus

Objective To investigate the correlation between proteolytic enzymes and microangiogenesis in degenerative intervertebral disc nucleus. Methods: Forty patients with degenerative disc nucleus pulposus who were admitted to our hospital were selected incase group and 20 healthy subjects were selected into the normal group. The specimens from the case group and the control group were collected to observe the degeneration of nucleus pulposus tissues with different degrees of classification, including H&E staining and immunohistochemical staining, and observe cathepsin such as aminopeptidase and vascular endothelial positive cells. The distribution of microvessels was also performed by the Weidner method. Results: After H&E staining, chondrocytes in the normal group clustered in the cartilage depression under the microscope. The matrix staining was uniform, while the number of chondrocytes in the case group decreased, and the nucleus was lightly stained or disappeared. Immunohistochemistry assay revealed little or no expression of aminopeptidase N (APN) and leucine aminopeptidase (LAP) in the nucleus pulposus of the normal group, but noticeable APN and LAP expressions in the degenerative intervertebral disc nucleus. Endothelial cells were stained singly or in clusters by CD31-labeled microvascular endothelial growth factor (VEGF). Conclusion: The expression level of various proteolytic enzymes such as aminopeptidase in the intervertebral disc and the linear relationship between microvessel formation and nerve fiber ingrowth in the intervertebral disc are helpful to further explore the molecular level of disc degeneration and pathophysiological mechanisms to aid clinical diagnosis and treatment of such diseases.

The establishment and evaluation of microvascular visualization model in bone tissue

Objective To explore a new method for visualization and morphologic study of intraosseous microvessels, measure the main parameters of the vesselsand evaluate the reliability of this model. Methods Abdominal aorta microfill perfusion in 10 New Zealand white rabbits were used. The specimens of the lower femur were fixed and decalcified sequentially, then the blood vessels were analyzed by microCT scan and three-dimensional reconstruction. The microvessels distribution and morphology of the bone were observed under the background of fluorescence by making the specimen into slices with a thickness of 300 μm. The main parameters of blood vessels were measured and evaluated. Results The structures of the intraosseous microvessels was clearly reconstructed in three dimensions by decalcification which can reduce the interference of the CT value of bone tissue to the CT value of the perfusion fluid. The distribution and morphology of the microvessels in the bone tissue were well displayed in the thick tissue slices in the fluorescence background imaging. There was no statistical difference in the number of blood vessels, mean vessel diameter and vascular percentage of the unit area between the results of microCT and histology (P=0.075, 0.921, 0.661). In histological Histology, the visual minimum blood vessel diameter reached the level of capillaries, significantly less than the results of microCT [(9.03±1.14) μm vs. (12.61±2.01) μm, P=0.038]. Conclusion MicroCT scan three-dimensional reconstruction combined with thick slice fluorescence background imaging can be used as a reliable method for the visualization and morphological study of intraosseous microvessels. Key words: Bone; Microvessel; Visualization; Histology; MicroCT

In Vivo Cell Tracking, Reactive Oxygen Species Scavenging, and Antioxidative Gene Down Regulation by Long-Term Exposure of Biomass-Derived Carbon Dots.

Biomass derived carbon dots (CD) have been observed to be excellent bioimaging probes due to their nontoxic, stable fluorescence, lesser bleachability, and excellent bioconjugation properties. In the current study, green chili extract derived CD synthesis via microwave irradiation is reported. The time dependent top down degradation of carbonaceous materials to CD are monitored via electron microscopy and correlated with fluorescence intensity. Further, the CD were explored for long-term cell tracking and cell therapy monitoring in a rodent model to study wound healing kinetics. The cells were monitorable up to 21 days (until the entire wound healed). CD were observed to scavenge reactive oxygen species (ROS) in vitro and in vivo and provided control over ROS scavenging enzyme gene expressions via down regulation. Further, it was observed to remodel the wound healing kinetics via altering granulation tissue distribution and formation of microvessels to establish the capability of CD to enhance wound healing.

Severe white matter astrocytopathy in CADASIL.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is characterized by strategic white matter (WM) hyperintensities on MRI. Pathological features include WM degeneration, arteriolosclerosis, lacunar infarcts, and the deposition of granular osmiophilic material. Based on the hypothesis that the gliovascular unit is compromised, we assessed the nature of astrocyte damage in the deep WM of CADASIL subjects. We evaluated post-mortem brains from CADASIL, cerebral small vessel disease, similar age cognitively normal and older control subjects. Standard immunohistochemical, immunofluorescent, and unbiased stereological methods were used to evaluate the distribution of astrocytes, microvessels, and autophagy markers in five different brain regions. Compared to the controls, the deep WM of CADASIL subjects overall showed increased numbers of glial fibrillary acidic protein (GFAP)-positive clasmatodendritic astrocytes (P=0.037) and a decrease in the percentage of normal appearing astrocytes (P=0.025). In accord with confluent WM hyperintensities, the anterior temporal pole contained abundant clasmatodendritic astrocytes with displaced aquaporin 4 immunoreactivity. Remarkably, we also found strong evidence for the immunolocalization of autophagy markers including microtubule-associated protein 1, light chain 3 (LC3), and sequestosome 1/p62 and Caspase-3 in GFAP-positive clasmatodendritic cells, particularly within perivascular regions of the deep WM. LC3 was co-localized in more than 90% of the GFAP-positive clasmatodendrocytes. Our novel findings show astrocytes undergo autophagy-like cell death in CADASIL, with the anterior temporal pole being highly vulnerable. We propose astrocytes transform from normal appearing type A to hypertrophic type B and eventually to clasmatodendritic type C cells. These observations also suggest the gliovascular unit of the deep WM is severely impaired in CADASIL.

Effectiveness of Superb Microvascular Imaging for the differentiation of intraductal breast lesions

The aim of this study was to compare the diagnostic performance of superb microvascular imaging (SMI) andpower Doppler imaging (PDI) for the differentiation of intraductal breast lesions. A total of 54 intraductalbreast lesions (39 benign, 10 atypical, and 5 malignant) in 53 patients were examined using B-mode ultrasonography,PDI, and SMI. Vascularity grading, distribution of microvessels, and penetrating vessels were evaluated using each Dopplertechnique. The diagnostic performances of both methods were compared. SMI was more efficient in detecting flowsignals than PDI (p=0.004). The highest diagnostic accuracy rates were achieved with SMI using vascular grading. Whenhypervascularity was used as a cut-off value to differentiate malignant and atypical lesions from benign lesions, the sensitivity,specificity, positive predictive value, negative predictive value, and accuracy were 66.6%, 80.7%, 66.6%, 80.7%, and 75.6%for PDI, and 86.6%, 76.9%, 68.4%, 90.9%, and 80.4% for SMI, respectively. SMI is more sensitive than PDI for detecting subtle blood flow in intraductal breast lesions with statistical significance. This novel and promising vascular imaging technique may be helpful in B-mode ultrasonography to distinguish intraductal breast lesions.