Endothelial CD31 Research Articles

Selective Internal Radiotherapy Alters the Profiles of Systemic Extracellular Vesicles in Hepatocellular Carcinoma.

Incidence of hepatocellular carcinoma (HCC) is increasing globally. Radioembolization (RE)/selective internal radiotherapy (SIRT) is a promising treatment for inoperable HCC. RE triggers an immune response, involving extracellular vesicles (EVs) which are crucial for cell communication and tumor development. This study explores EV immune profiles and origins in patients with inoperable HCC before and after SIRT/RE. Blood samples from 50 HCC-patients treated with SIRT/RE were collected before and after therapy to determine cytokines and isolate EVs using size exclusion chromatography. The dynamic range and EV quality required for detecting variations in surface markers were assessed. Thirty-seven EV surface markers were analyzed using flow cytometry and correlated with clinical parameters. Several immunological markers (CD4, CD2, CD40, CD45, CD49e, CD69, CD209-EVs) were present in the circulation of HCC patients. These markers positively correlated with therapy response and survival. Conversely, B cell CD20, endothelial cell CD146, platelet CD49e, and CD41b EV markers negatively correlated with 60-day survival. Elevated levels of IL-6 and IL-8 before therapy correlated negatively with patient survival, coinciding with a positive correlation with CD20-positive EVs. Plasma EVs from HCC patients exhibit immunological, cancer, and coagulation markers, including potential biomarkers (CD4, CD20, CD49e, CD146). These may enhance our understanding of cancer biology and facilitate SIRT therapy monitoring.

Comparison the therapeutic effects of bone marrow CD144+ endothelial cells and CD146+ mesenchymal stem cells in POF rats.

Premature ovarian insufficiency (POI) is a challenging issue in terms of reproduction biology. In this study, therapeutic properties of bone marrow CD146+ mesenchymal stem cells (MSCs) and CD144+ endothelial cells (ECs) were separately investigated in rats with POI. POI rats were classified into control POI, POI + CD146+ MSCs, and POI + CD144+ ECs groups. Enriched CD146+ MSCs and CD144+ ECs were directly injected into ovarian tissue (15 × 104 cells/10 μL) in relevant groups. After 4 weeks, follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) levels were measured in blood samples. Ovarian tissues were collected and subjected to Hematoxylin-Eosin and Masson's trichrome staining. The expression of angp-2, vegfr-2, smad-2, -4, -6, and tgf-β1 was studied using qRT-PCR analysis. Histopathological examination indicated an increased pattern of atretic follicles in the POI group related to the control rats (P<0.0001). Data indicated that injection of POI + CD146+ MSCs and CD144+ ECs in POI rats reduced atretic follicles and increased the number of normal follicles (P<0.01). Along with these changes, the content of blue-colored collagen fibers was diminished after cell transplantation. Besides, cell transplantation in POI rats had the potential to reduce increased FSH, and LH levels (P<0.05). In contrast, E2 content was increased in POI + CD146+ MSCs and POI + CD144+ ECs groups compared to control POI rats, indicating restoration of follicular function. CD144+ (smad-2, and -4) and CD146+ (smad-6) cells altered the activity of genes belonging TGF-β signaling pathway. Unlike POI + CD146+ MSCs, aberrant angiogenesis properties were significantly down-regulated in POI + CD144+ ECs related to the control POI group (P<0.05). The transplantation of bone marrow CD146+ and CD144+ cells can lead to the restoration of ovarian tissue function in POI rats via modulating different mechanisms associated with angiogenesis and fibrosis.

Naru-3 inhibits inflammation, synovial hyperplasia, and neovascularization in collagen-induced arthritis in rats.

Naru-3 is a prescribed formulation based on the theory of Mongolian medicine for the treatment of rheumatoid arthritis (RA). Naru-3 consists of three medicinal agents: Aconitum kusnezoffii Reichb (caowu), Terminalia chebula Retz (hezi), and Piper longum L (biba). These medicinal agents are widely distributed in the Mongolian area of China and have been used to treat rheumatism for centuries. Mongolian medicine Naru-3 is commonly prescribed to treat RA, but its mechanism of action is unknown. A rat collagen-induced arthritis (CIA) model was established to investigate the mechanism of Naru-3. Rats were treated with Naru-3, Etanercept (ETN), and sodium carboxymethylcellulose (CMC) for four weeks. After treatment was terminated, paw thickness, ankle diameter, and arthritis index (AI) were scored. Synovial hyperplasia was evaluated using hematoxylin and eosin (H&E) staining and two-dimensional ultrasonography. Synovitis and neovascularization were assayed using power Doppler imaging (PDI) and contrast-enhanced ultrasonography (CEUS). Levels of vascular endothelial growth factor (VEGF), interleukin (IL)-1, and CD31 in the serum or synovium were detected using ELISA and immunohistochemistry analyses. Naru-3 and ETN alleviated the symptoms of CIA as evidenced by diminished paw thickness, ankle diameter, and AI scores. Mechanistically, Naru-3 inhibited synovial hyperplasia, synovitis, and neovascularization by diminishing systemic and local inflammation, as indicated by the relative expression of VEGF and IL-1 in the serum and synovium. After four weeks of treatment, no significant neovascularization was observed in the Naru-3 group, but neovascularization and synovitis occurred in the ETN group, as demonstrated by H&E staining, PDI, and CEUS examination. Naru-3 inhibited inflammation, synovial hyperplasia, and neovascularization and alleviates RA in our CIA rat model. No symptom recurrence was observed four weeks after drug treatment.

Biomarkers of lung alveolarization and microvascular maturation in response to intermittent hypoxia and/or early antioxidant/fish oil supplementation in neonatal rats.

Extremely preterm infants experience frequent intermittent hypoxia (IH) episodes during oxygen therapy which causes significant damage to the lungs and curtails important signaling pathways that regulate normal lung alveolarization and microvascular maturation. We tested the hypothesis that early supplementation with fish oil and/or antioxidants in rats exposed to neonatal IH improves expression of lung biomarkers of alveolarization and microvascular maturation, and reduces IH-induced lung injury. From birth (P0) to P14, rat pups were exposed to room air (RA) or neonatal IH during which they received daily oral supplementation with either: (1) olive oil (OO) (control); (2) Coenzyme Q10 (CoQ10) in OO; (3) fish oil; (4) glutathione nanoparticles (nGSH); or (5) fish oil +CoQ10. At P14 pups were placed in RAuntil P21 with no further treatment. RA controls were similarly treated. Lung growth and alveolarization, histopathology, apoptosis, oxidative stress and biomarkers of alveolarization and microvascular maturation were determined. Neonatal IH was associated with reduced lung weights and severe histopathological outcomes. These effects were curtailed with fish oil and nGSH. nGSH was also protective against apoptosis, while CoQ10 prevented IH-induced ROS production. Of all treatments, nGSH and CoQ10 + fish oil-induced vascular endothelial growth factor165 and CD31 (Platelet endothelial cell adhesion molecule-1), which are associated with angiogenesis. CoQ10 + fish oil improved alveolarization in RA and IH despite evidence of hemorrhage. The benefits of nGSH and CoQ10 + fish oil suggest an antioxidant effect which may be required to curtail IH-induced lung injury. Further clinical assessment of the effectiveness of nGSH is warranted.

Endothelial Cd36 mediates endothelium stiffening and promotes barrier disruption in high-fat diet fed mice

The impact of a Western-style high fat diet (HFD) on cardiovascular health is a chronic concern. Our previous studies showed that HFD results in significant endothelial cell (EC) stiffening, which was abrogated by the global deletion of scavenger receptor Cd36. Here we present studies into the role of endothelial Cd36 on EC stiffening and EC permeability in mice fed a short-term HFD of 6-8 weeks. Inducible EC-specific knockdown of Cd36 (EC-Cd36-KD) mice were generated by crossing EC-promoter Cre mice with Cd36fl/fl mice. Knockdowns were induced via tamoxifen injections at 8 weeks of age, after which mice were placed on HFD for 6-8 weeks (Cre-only and sham injected mice served as control groups). The efficiency and specificity of EC-Cd36-KD was verified through RT-qPCR and immunostaining. At the end of the study window, control and EC-Cd36-KD mice exhibited hallmarks of metabolic disease like significant increases in body weight and blood cholesterol (total and LDL-Chol). EC-Cd36-KD did not have an effect on these parameters. As described before, HFD results in significant endothelial stiffening in freshly-isolated intact aortas, assessed using atomic force microscopy. We show here that EC-Cd36-KD prevented HFD-induced EC stiffening. No effect was observed in EC-Cd36-KD mice fed low fat diet. In contrast no change was observed in vascular wall stiffness, assessed via pulse wave velocity analysis following Doppler echocardiography in the ventral short-axis. The EC stiffening effect was observed only in male mice, with female mice showing no EC stiffening and exhibiting lower levels of weight gain and blood cholesterol in comparison to males. EC permeability of the aorta was assessed by injecting Evans blue dye (EBD) into circulation and measuring its penetration into the vascular wall by fluorescent imaging. While HFD results in increased EC permeability, EC-Cd36-KD significantly reduced the penetration of EBD into the aortic wall indicating a protective barrier effect. Cre-only HFD-fed controls were unaffected. These results suggest that Cd36-depedent EC stiffening mediates EC permeability of the aorta. Mechanistically, we show that Cd36 is required for oxLDL-induced stiffening but not for the EC stiffening induced by 7-ketocholesterol and oxidized phospholipid PGPC, suggesting that these oxidized lipids induce endothelial stiffening downstream of Cd36. Finally, the knockdown of RhoGDI-1, an inactivator of RhoA, using siRNA prevents oxysterol-induced EC stiffening. We propose that oxidized lipids induce EC stiffening by uncoupling of RhoA from RhoGDI-1 leading to its activation. This work was supported by NHLBI grants HL141120, HL073965, HL083298 (IL), NHLBI trainee award T32HL144459 and an internal fellowship (VA). This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Upregulation of Cell Surface Glycoproteins in Correlation with KSHV LANA in the Kaposi Sarcoma Tumor Microenvironment.

HIV-associated epidemic Kaposi sarcoma (EpKS) remains one of the most prevalent cancers in sub-Saharan Africa despite the widespread uptake of anti-retroviral therapy and HIV-1 suppression. In an effort to define potential therapeutic targets against KS tumors, we analyzed previously published KS bulk tumor transcriptomics to identify cell surface biomarkers. In addition to upregulated gene expression (>6-fold) in the EpKS tumor microenvironment, biomarkers were selected for correlation with KSHV latency-associated nuclear antigen (LANA) expression. The cell surface glycoprotein genes identified were KDR, FLT4, ADAM12, UNC5A, ZP2, and OX40, as well as the endothelial lineage determinants Prox-1 and CD34. Each protein was evaluated for its expression and co-localization with KSHV LANA using multi-color immunofluorescence in KS tissues, KSHV-infected L1T2 cells, uninfected TIVE cells, and murine L1T2 tumor xenografts. Five surface glycoproteins (KDR, FLT4, UNC5A, ADAM12, and CD34) were associated with LANA-positive cells but were also detected in uninfected cells in the KS microenvironment. In vitro L1T2 cultures showed evidence of only FLT4, KDR, and UNC5A, whereas mouse L1T2 xenografts recapitulated human KS cell surface expression profiles, with the exception of CD34 and Prox-1. In KS tumors, most LANA-positive cells co-expressed markers of vascular as well as lymphatic endothelial lineages, suggesting KS-associated dedifferentiation to a more mesenchymal/progenitor phenotype.

Characterization of the Tumor Microenvironment in Jaw Osteosarcomas, towards Prognostic Markers and New Therapeutic Targets

Background-The purpose of this study was to investigate the bone resorption, as well as the vascular and immune microenvironment, of jaw osteosarcomas (JO) and to correlate these features with patient clinical outcomes. Methods-We studied 50 JO biopsy samples by immunohistochemical analysis of tissue microarrays (TMAs). We investigated the bone remodeling markers RANK/RANKL/OPG, the endothelial glycoprotein CD146, and biomarkers of the immune environment (CD163 and CD68 of macrophages, CD4+ and CD8+ of tumor-infiltrating lymphocytes (TILs), and an immune checkpoint PD-1/PD-L1). The biomarkers were analyzed for their influence on progression (recurrence and metastasis), overall survival (OS), and disease-free survival (DFS). Results-A strong and significant correlation has been found between CD163 staining and lower OS and DFS. The level of CD4+ and CD8+ staining was low and non-significantly associated with survival outcomes. High levels of RANK and RANKL were found in the tumor samples and correlated with lower DFS. Conclusion-Our findings suggest that CD163+ TAMs represent markers of poor prognosis in JO. Targeting TAMs could represent a valuable therapeutic strategy in JO.

Correlation of Microvessel Density with Clinicothological Features of Hodgkin Lymphoma

BackgroundAngiogenesis is the process of new blood vessel growth that has an important role in tumor development. Microvessel density (MVD) assessment is one of the methods to assess angiogenesis. A high MVD is associated with a poor prognosis in various types of cancer including Hodgkin lymphoma (HL). Although, HL incidence is lower in developing countries, the death rate is higher in these regions. Moreover, this type of lymphoma is higher among young and reproductive people. The aim of this study was to determine the association between microvessel density and HL clinicopathological characteristics.MethodsThis was an observational analytic study with a cross-sectional method, conducted at the Anatomical Pathology Department/Dr. Mohammad Hoesin Palembang. The samples were paraffin blocks diagnosed as HL for period 2016-2021 and had complete clinicopathological data according to the criteria of this study. The slides were immunostained with antibody anti-CD34. One microvessel was determined by observing the endothelial morphology and CD34 expression in endothelial cytoplasm and/or cell membrane, then was evaluated and calculated by Image J. Data was statistically analyzed by STATA version 15.ResultsMajority HLs in this study were discovered in the age group of &lt;45 years (71.00%), male (61.30%), nodal location (80.60%) and lymphocyte rich classical HL (LRCHL) subtype (51.60%). Although there was no significant association between MVD and the clinicopathological characteristics of HL, high MVD tend to occur in the age &lt;45 years, male, nodal location, and LRCHL subtype.ConclusionThere was no significant association between microvessel density and HL clinicopathological characteristics.

Effect of Moxibustion Combined With Cisplatin on Tumor Microenvironment Hypoxia and Vascular Normalization in Lewis Lung Cancer Mice.

This study was developed to evaluate the effects of moxibustion on tumor microenvironmental hypoxia in a murine model of Lewis lung carcinoma (LLC). Twenty-four tumor-bearing mice were randomized into tumor group (T), tumor + cisplatin group (TC), tumor + moxibustion group (TM), and tumor + cisplatin + moxibustion group (TMC) (n = 6/group). Six age-matched C57BL/6 mice were employed as control group (Ctrl). A tumor model was established by implanting LLC cells into the right flank of each mouse. Animals in the TM group received moxibustion treatment at the ST36 (bilateral) and GV4 acupoints on the day of visible tumor formation. Moxibustion treatment was performed every other day for a total of 7 sessions. Animals in the TC group were intraperitoneally injected with cisplatin (3 mg/kg) on day 3 after visible tumor formation, and this treatment was performed every 3 days for 4 times. Animals in the TMC group underwent combined moxibustion and chemotherapy treatment, following the same conditions as outlined above. Following treatment, the concentrations of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), CD31, and Ki67 were measured using ELISA, Western blot, and immunohistochemical staining. Compared to the tumor group, treatment in the TM, TC, and TCM groups resulted in varying reductions in tumor growth (P < .001 or P < .05), while tumor microenvironmental hypoxia was alleviated as evidenced by the downregulation of HIF-1α, VEGFA, and CD31(P < .001-P < .05). Our results suggest that a combined approach of moxibustion and cisplatin can alleviate intratumoral hypoxia, promote vascular normalization, and slow the growth of LLC tumors in mice.

Low-energy shock wave therapy ameliorates ischemic-induced overactive bladder in a rat model

This study was to evaluate whether Low-energy shock wave therapy (LESW) improves ischemic-induced overactive bladder in rats and investigate its therapeutic mechanisms. Sixteen-week-old male Sprague–Dawley rats were divided into three groups: arterial injury (AI), AI with LESW (AI-SW), and control groups. LESW was irradiated in AI-SW during 20–23 weeks of age. At 24 weeks of age, conscious cystometry was performed (each n = 8). The voiding interval was shortened in AI (mean ± SEM: 5.1 ± 0.8 min) than in control (17.3 ± 3.0 min), whereas significant improvements were observed in AI-SW (14.9 ± 3.3 min). The bladder blood flow was significantly increased in AI-SW than in AI. Microarray analysis revealed higher gene expression of soluble guanylate cyclase (sGC) α1 and β1 in the bladder of AI-SW compared to AI. Protein expression of sGCα1 and sGCβ1 was higher in AI-SW and control groups than in AI. Cyclic guanosine monophosphate (cGMP) was elevated in AI-SW. As an early genetic response, vascular endothelial growth factor and CD31 were highly expressed 24 h after the first LESW. Suburothelial thinning observed in AI was restored in AI-SW. Activation of sGC-cGMP may play a therapeutic role of LESW in the functional recovery of the bladder.

Abstract 11511: Stem Cell-Derived Exosome Nebulization Therapy (SCENT) Promotes Cardiac Repair After Myocardial Infarction by Inhibiting CD36

Introduction: Stem cell-derived exosomes are promising therapeutics for cardiac repair after injury. Since they have short half-life in vivo, efficient exosome therapies need repeated dosing. Here, we develop a noninvasive and repeatable method for exosome delivery via a nebulizer after myocardial infarction (MI), named Stem Cell-Derived Exosome Nebulization Therapy (SCENT). Methods: Lung spheroid cell-derived-exosomes were isolated from secretome by ultrafiltration and characterized by a Zetasizer. We labelled them with a NIR-680 dye, before delivering them to C57BL/6 mice with MI model via a nebulizer for 7 consecutive days. IVIS live imaging and fluorescent microscopy was used to detect the biodistribution of exosomes in vivo and ex vivo. Echocardiographies were performed to monitor the cardiac function after SCENT in MI mouse and histological analysis helped investigating the myocardial repair, in comparison with the controls. Single-cell sequencing of the whole heart was performed to explore the mechanism of action after SCENT. Results: Homing of exosomes to the ischemic heart after SCENT was detected by immunofluorescence microscopy (Fig. 1a). SCENT led to significantly improved left ventricular function to our mouse MI model at 2 weeks (EF: 66.97 ± 2.49%), compared to that of MI control (EF: 36.13 ± 3.57%) (Fig. 1b) , along with reduced infarct size (Fig. 1c). Single-cell sequencing revealed a significant downregulation of CD36 in cardiac endothelial cells, which suppressed the transport and uptake of fatty acids in myocardium, leading to a compensatory increase in glucose utilization and cardiomyocyte proliferation (Fig. 1d,e) . Conclusions: Exosomes delivered by SCENT were recruited to the ischemic heart, which promoted function recovery and cardiac repair in a mouse MI model. This therapeutic effect was attributed to the inhibition of endothelial CD36. SCENT gives a promising solution for the repeatable and noninvasive treatment for heart repair.

Skin endothelial cell and microcirculation function study in recurred keloids patients after keloid surgery and radiotherapy.

Keloid is a type of benign tumor of the skin with abnormal proliferation of fibrous tissue. We sought to observe the changes in skin microcirculation and endothelial cell function around the recurred keloid and explore the skin microcirculation characters in recurred keloid patients. Six patients with recurred keloid were treated with keloid surgery and radiotherapy for the second time. Microcirculation of recurred keloids and their surrounding normal skin tissue was observed with laser Doppler flowmeter before operation. Expression of vascular endothelial growth factor (VEGF), CD31, and HIF-1α were identified by several assay. The local blood flow of group RN was enhanced. The average strength of group N is 0.87. The average strength of group RN is 2.08. The expression of VEGF, CD31, and hypoxia inducible factor-1α (HIF-1α) protein in the keloid-recurred skin (RN) group was higher than the normal skin group via immunohistochemistry (IHC) and Western blotting analysis. The relative expression of VEGF and CD31 mRNA was significantly increased in RN group samples (P < .05). There are significant differences in the expression of VEGF, CD31, and HIF-1α in the recurred keloid skin after radiotherapy and normal skin. They may be used as potential biomarkers and targets for future research on keloid recurrence.

Taurine-Upregulated Gene 1 Attenuates Cerebral Angiogenesis following Ischemic Stroke in Rats.

Angiogenesis is one of the therapeutic targets of cerebral infarction. Long noncoding RNAs (lncRNAs) can regulate the pathological process of angiogenesis following ischemic stroke. Taurine-upregulated gene 1 (TUG1), an lncRNA, is correlated to ischemic stroke. We intended to determine the effect of TUG1 on angiogenesis following an ischemic stroke. Middle cerebral artery occlusion (MCAO) was adopted to build a focal ischemic model of the rat brain, and pcDNA-TUG1 and miR-26a mimics were injected into rats. Neurological function was estimated through modified neurological severity scores. The volume of focal brain infarction was calculated through 2,3,5-triphenyltetrazolium chloride staining. The level of TUG1 and miR-26a was measured by PCR. The expression of vascular endothelial growth factor (VEGF) and CD31 was checked using immunohistochemistry and western blot. The correlation between miR-26a and TUG1 was verified through a luciferase reporter assay. TUG1 increased noticeably while miR-26a was markedly reduced in MCAO rats. Overexpression of miR-26a improved neurological function recovery and enhanced cerebral angiogenesis in MCAO rats. TUG1 overexpression aggravated neurological deficits and suppressed cerebral angiogenesis in MCAO rats. Bioinformatics analysis revealed that miR-26a was one of the predicted targets of TUG1. Furthermore, TUG1 combined with miR-26a to regulate angiogenesis. TUG1 overexpression antagonized the role of miR-26a in neurological recovery and angiogenesis in MCAO rats. TUG1/miR-26a, which may act as a regulatory axis in angiogenesis following ischemic stroke, can be considered a potential target for cerebral infarction therapy.

Macrophage Phenotypes and Gene Expression Patterns Are Unique in Naturally Occurring Metabolically Healthy Obesity.

Obesity impacts 650 million individuals globally, often co-occurring with metabolic syndrome. Though many obese individuals experience metabolic abnormalities (metabolically unhealthy obese [MUO]), ~30% do not (metabolically healthy obese [MHO]). Conversely, >10% of lean individuals are metabolically unhealthy (MUL). To evaluate the physiologic drivers of these phenotypes, a 44-animal African green monkey cohort was selected using metabolic syndrome risk criteria to represent these four clinically defined health groups. Body composition imaging and subcutaneous adipose tissue (SQ AT) biopsies were collected. Differences in adipocyte size, macrophage subtype distribution, gene expression, vascularity and fibrosis were analyzed using digital immunohistopathology, unbiased RNA-seq, endothelial CD31, and Masson’s trichrome staining, respectively. MHO AT demonstrated significant increases in M2 macrophages (p = 0.02) and upregulation of fatty acid oxidation-related terms and transcripts, including FABP7 (p = 0.01). MUO AT demonstrated downregulation of these factors and co-occurring upregulation of immune responses. These changes occurred without differences in AT distributions, adipocyte size, AT endothelial cells, collagen I deposition, or circulating cytokine levels. Without unhealthy diet consumption, healthy obesity is defined by an increased SQ AT M2/M1 macrophage ratio and lipid handling gene expression. We highlight M2 macrophages and fatty acid oxidation as targets for improving metabolic health with obesity.

Management of wounds with exposed bone structures using an induced-membrane followed by polymethyl methacrylate and second-stage skin grafting in the elderly with a 3-year follow-up.

The treatment of traumatic wounds with exposed bone or tendons is often challenging. An induced membrane (IM) is used to reconstruct bone defects, as it provides an effective and sufficient blood supply for bone and soft-tissue reconstruction. This study explored a novel two-stage strategy for wound management, consisting of initial wound coverage with polymethyl methacrylate (PMMA) and an autologous split-thickness skin graft under the IM. Fifty inpatients were enrolled from December 2016 to December 2019. Each patient underwent reconstruction according to a two-stage process. In the first stage, the defect area was thoroughly debrided, and the freshly treated wound was then covered using PMMA cement. After 4-6 weeks, during the second stage, the PMMA cement was removed to reveal an IM covering the exposed bone and tendon. An autologous split-thickness skin graft was then performed. Haematoxylin and eosin (H&E) staining and immunohistochemical analysis of vascular endothelial growth factor (VEGF), CD31 and CD34 were used to evaluate the IM and compare it with the normal periosteal membrane (PM). The psychological status and the Lower Extremity Function Scale (LEFS) as well as any complications were recorded at follow-up. We found that all skin grafts survived and evidenced no necrosis or infection. H&E staining revealed vascularised tissue in the IM, and immunohistochemistry showed a larger number of VEGF-, CD31- and CD34-positive cells in the IM than in the normal PM. The duration of healing in the group was 5.40 ± 1.32 months with a mean number of debridement procedures of 1.92 ± 0.60. There were two patients with reulceration in the group. The self-rating anxiety scale scores ranged from 35 to 60 (mean 48.02 ± 8.12). Postoperatively, the LEFS score was 50.10 ± 9.77. Finally, our strategy for the management of a non-healing wound in the lower extremities, consisting of an IM in combination with skin grafting, was effective, especially in cases in which bony structures were exposed in the elderly. The morbidity rate was low.

Angiosarcoma of the visceral organs: A morphological, immunohistochemical, and C-MYC status analysis.

Visceral angiosarcomas are rare malignant tumors with poor prognosis. The pathogenesis remains unclear. Herein, we describe the morphological, immunohistochemical, and C-MYC status of a series of visceral angiosarcomas. We evaluated the clinicopathologic and C-MYC status of visceral angiosarcomas (n=12) and compared them to a control series of angiosarcomas arising in cutaneous (n=15) and soft tissue structures (n=15). Clinical follow-up data were obtained for all patients and exhibited high metastasis and mortality rates. Malignant endothelial cells displayed a range of morphological features including nonepithelioid, epithelioid, and mixed features, forming vasoformative (n=6), solid (n=4) or mixed (n=2) architectures. Epithelioid morphology was present in 4/12 tumors. Mitoses ranged from 3 to 60 per 10 high-power fields. Necrosis was observed in 10/12 tumors. By immunohistochemistry, all angiosarcomas expressed at least 2 markers of endothelial differentiation, including CD31, CD34, vWF, ERG, and Fli-1. Eight cases of C-MYC amplification and 5 cases of C-MYC translocation were detected. Our data showed that visceral angiosarcoma is more common in women, and the clinical presentations of patient age and tumor size were significantly different between the study and control groups. No significant difference in staining between the visceral angiosarcoma and control groups was observed for endothelial markers, while different C-MYC statuses were detected.

Морфологічні та імуногістохімічні особливості плацент у породіль з інфекціями, що передаються статевим шляхом, в анамнезі

Purpose - to explore and evaluate macroscopic, histological, morphological and immunohistochemical changes in the placenta, to study the peculiarities of the marker expression of endothelial cells CD34 in the vascular bed of the placental villous tree in women in labour with a history of sexually transmitted infections (STIs) after pregravid preparation before cycles of assisted reproductive technologies (ART). Materials and methods. The analysis of morphological and immunohistochemical examination as well as expression level of CD34 in 50 placentas from women in labour with a history of STIs was conducted. The Group I (main) - 25 placentas from the women in labour who underwent pregravid preparation before the ART cycle, obstetric and perinatal support and delivery in accordance with the medical and organizational algorithms developed by us, prognostic methods and treatment and preventive regimens; the Group II - 25 placentas from the women in labour who received generally accepted prognostic and therapeutic and preventive measures. Results. Histological examination of placentas from the women of the Group I demonstrated the manifestations of compensatory processes prevailed, and there was a much lower severity of pathological changes. The placental coefficient in the women of the Group I was probably higher than in women of the Group II, accounting for 0.17 versus 0.15. There was a decrease in the branching of blood vessels, as a result of which the capillaries occupied mainly the central and paracentral position. Dystrophic and necrotic processes, with the replacement of the chorion epithelium with fibroid masses, were manifested in a small number of terminal villi which belonged to the adaptive mechanisms. The largest area of CD34 expression in the villous chorion of the placenta was observed in the Group I and was 9.49±0.47%, in the Group II it was 1.29 times lower (7.34±0.15%; p&lt;0.01). The highest optical density of CD34 expression in the villi chorion of the placenta was observed in the women of the Group II (0.22±0.01 r.u.), which was 1.25 times higher than in patients of the Group I (0.20±0.01 r.u.; p&lt;0.01). Conclusions. In the case of pregravid preparation before ART cycles and in the case of complex correction of maladaptive disorders in the fetoplacental complex of pregnant women with a history of STIs, all structural mechanisms of placental adaptation are included, which allow to maintain the morphometric and diffusion parameters of the villous tree at the level of stable compensation, which is the most important adaptive tool that helps to maintain fetal viability. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the author.

Involvement of endothelial progenitor cells in blood flow recovery through activation of the Wnt/β-catenin signaling pathway and inhibition of high oxidative stress in diabetic hindlimb ischemic rats.

Diabetes mellitus (DM) often causes stenosis and occlusion of hindlimb blood vessels, which are also the main cause for hindlimb ischemia in elderly people. To investigate the therapeutic effect of endothelial progenitor cell (EPC) transplantation on diabetic hindlimb ischemia. Endothelial progenitor cells were separated, labeled with PKH-26 and transplanted into rat models (107 cells/100 g). Dichlorodihydrofluorescein diacetate (DCFH-DA) was used to detect any oxidative stress. Streptozotocin (STZ) was injected to establish a diabetic rat model and hindlimb ischemia model was established via operation. Western blotting was used to detect total β-catenin (T-β-catenin) and non-phospho-β-catenin (NP-β-catenin) levels. The malondialdehyde (MDA), superoxide dismutase (SOD), Wnt3a, Wnt5a and Wnt7a levels were detected using enzyme-linked immunosorbent assay (ELISA). Oxidative stress was measured using DCFH-DA and dihydroethidium (DHE). The endothelial biomarker CD31 was observed to highlight vessels, and PKH-26 to trace migration/adhesion of EPCs. Endothelial progenitor cells were successfully isolated and identified, and diabetic hindlimb ischemic rat models were created. Tempol remarkably improved blood flow in diabetic hindlimb ischemic rats compared to DM+EPCs rats at 14 days (p < 0.001) and 28 days post-operation (p < 0.001). High oxidative stress was observed in diabetic hindlimb ischemic rats. Tempol significantly inhibited oxidative stress levels in diabetic hindlimb ischemic rats. Furthermore, Tempol significantly promoted angiogenesis in diabetic hindlimb ischemic rats compared to DM+EPCs rats. The β-catenin inhibitor, XAV (DM+EPCs+Tempol+XAV group), significantly suppressed blood flow recovery and angiogenesis in diabetic hindlimb ischemic rats when compared to the DM+EPCs+Tempol group at 14 days (p = 0.026) and 28 days (p < 0.001). The XAV remarkably reduced T-β-catenin (p < 0.001) and N-β-catenin (p = 0.030) levels in Tempol-treated diabetic hindlimb ischemic rats, as compared to the DM+EPCs+Tempol group. The Wnt5a participated in the pathology of diabetic hindlimb ischemia. There are high oxidative stress levels in both EPCs in high-glucose environments and diabetic hindlimb ischemia, which can lead to limited blood flow recovery. The high oxidative stress caused the inhibition of Wnt/β-catenin signaling pathway, leading to limited blood flow recovery in diabetic hindlimb ischemia. At the same time, Wnt5a participated in the EPC-mediated blood flow recovery.

Effect of platelet concentrate prepared by different methods on the healing of full-thickness skin defects.

Platelet-rich plasma and concentrated growth factors have been widely utilized in tissue regeneration. However, very few studies have focused on comparing the merits of these two materials in skin repair. We aim to compare the wound healing effects of four platelet concentrates. Auto-platelet-rich plasma, artificial platelet-rich plasma, concentrated growth factors in a liquid phase, and a gel phase were prepared, and a full-thickness skin defect model of mice was made. The skin defects were treated with normal saline as a control and also the four kinds of platelet concentrates mentioned above, respectively. Wound size was measured and calculated on days 3, 5, 7, and 10, with histological analysis performed. All four platelet concentrates accelerated wound healing in mice. Interleukin-1β (IL-1β) and tumor necrosis factor (TNF-α) in concentrated growth factors in a liquid phase, a gel phase, and artificial platelet-rich plasma groups were significantly lower than those in the control group; and vascular endothelial growth factor (VEGF) and CD34 were significantly higher than those in the control group and auto-platelet-rich plasma group. All four platelet concentrates appear to promote wound healing. Compared with auto-platelet-rich plasma, concentrated growth factors in a liquid phase and in a gel phase, and artificial platelet-rich plasma seem to have more substantial effects in promoting angiogenesis, epithelialization, and reducing inflammation, thus promoting a stronger effect on wound healing.

Targeting Hydrogen Sulfide Modulates Dexamethasone-Induced Muscle Atrophy and Microvascular Rarefaction, through Inhibition of NOX4 and Induction of MGF, M2 Macrophages and Endothelial Progenitors.

Long-term use of Glucocorticoids produces skeletal muscle atrophy and microvascular rarefaction. Hydrogen sulfide (H2S) has a potential role in skeletal muscle regeneration. However, the mechanisms still need to be elucidated. This is the first study to explore the effect of Sodium hydrosulfide (NaHS) H2S donor, against Dexamethasone (Dex)-induced soleus muscle atrophy and microvascular rarefaction and on muscle endothelial progenitors and M2 macrophages. Rats received either; saline, Dex (0.6 mg/Kg/day), Dex + NaHS (5 mg/Kg/day), or Dex + Aminooxyacetic acid (AOAA), a blocker of H2S (10 mg/Kg/day) for two weeks. The soleus muscle was examined for contractile properties. mRNA expression for Myostatin, Mechano-growth factor (MGF) and NADPH oxidase (NOX4), HE staining, and immunohistochemical staining for caspase-3, CD34 (Endothelial progenitor marker), vascular endothelial growth factor (VEGF), CD31 (endothelial marker), and CD163 (M2 macrophage marker) was performed. NaHS could improve the contractile properties and decrease oxidative stress, muscle atrophy, and the expression of NOX4, caspase-3, Myostatin, VEGF, and CD31 and could increase the capillary density and expression of MGF with a significant increase in expression of CD34 and CD163 as compared to Dex group. However, AOAA worsened the studied parameters. Therefore, H2S can be a promising target to attenuate muscle atrophy and microvascular rarefaction.