Collagen Fibers In Tissue Research Articles

Integrated pathological, proteomic and metabolomic analyses reveal significant changes of Eriocheir sinensis hepatopancreatic in response to the microsporidian Hepatospora eriocheir infection

Infection by Hepatospora eriocheir would cause serious disease, hepatopancreatic necrosis disease (HPND), and lead to significant economic losses in Eriocheir sinensis. In this study, the response of E. sinensis during H. eriocheir infection was explored by PCR detection, physiological determination (Masson staining and Calcofluor White staining), electron microscopy, and integrated proteome and metabolome analyses. Pathological and electron microscopy analysis demonstrated that H. eriocheir infection could severely damage the E. sinensis hepatopancreas tissue collagen fibers, muscle fibers, and the cells ultrastructure. Base on the PCR detection and Calcofluor White staining results selected the H. eriocheir positive or negative E. sinensis hepatopancreas used to proteome and metabolome analyses. Using a 2-fold change as a physiologically significant benchmark, total of 370 significantly changed proteins (225 up-regulated proteins and 145 down-regulated proteins) and 386 significantly different metabolites (217 metabolites up-regulated and 169 metabolites down-regulated) in crab hepatopancreas during H. eriocheir infection were identified by proteomic and metabolomic analysis, respectively. Biological analysis of these significantly changed proteins and metabolites showed that the lipid metabolism (for example, glycerophospholipid metabolism, carnitine metabolism, lysolecithin metabolism, biosynthesis of unsaturated fatty acids, sphingolipid metabolism, and linoleic acid metabolism) and energy metabolism (such as tricarboxylic acid cycle (TCA cycle), oxidative phosphorylation, and ATP-binding cassette transporter (ABC transporters)) of E. sinensis hepatopancreas were disproportionally affected by H. eriocheir infection. Meanwhile, several key immune-related processes, proteins or metabolites were activated to against the H. eriocheir infection, for example apoptosis, phagocytosis, oxidative stress, C-type lection, β-1, 3-glucan binding protein, vitellogenin receptor, arginine, leucine, aspartic acid, lysine. This study could provide an insightful view into the response mechanism in E. sinensis hepatopancreas during H. eriocheir infection.

Icariin alleviates ferroptosis-related atherosclerosis by promoting autophagy in xo-LDL-induced vascular endothelial cell injury and atherosclerotic mice.

Vascular endothelial cells (VECs) are located between the blood plasma and the vascular tissue, and the ferroptosis (iron-dependent programmed cell death) of VECs can lead to a range of cardiovascular diseases. Icariin is the main active ingredient of Epimedium brevicornum Maxim., which can improve endothelial cell dysfunction. In the present study, the protective effects of icariin on oxidised low-density lipoprotein (ox-LDL)-treated VECs and high-fat diet-fed Apolipoprotein E-deficient mice were investigated. Inflammatory fibrosis in tissues and inflammatory factors in serum and cell supernatants were detected, and mitochondrial membrane potential and the expression levels of ferroptosis-associated proteins were also detected. The results revealed that icariin reduced the endothelial atherosclerotic plaque area and collagen fibres in aortic sinus tissue, and increased the viability and mitochondrial membrane potential, whereas it reduced the reactive oxygen species levels of VECs. The nucleation of transcription factor EB (TFEB) and subsequent autophagy were negatively associated with ferroptosis in endothelial cells, and the more prominent the autophagy, the lower the levels of ferroptosis. Furthermore, by co-treating the cells with icariin and the two autophagy inhibitors, Bafilomycin A1 (blocking autophagosome and lysosome fusion) and 3-methyladenine (blocking autophagosome formation), respectively, the promoting effects of icariin on autophagy were found to be mediated through the process of autophagosome-lysosome fusion. In in vivo experiments, icariin reduced ferroptosis, alleviated atherosclerotic lesions and increased the rate of TFEB nucleation. Additionally, it was found that ARG304, THR308 and GLN311 were the optimal binding sites for the interaction between icariin and TFEB. Taken together, these results suggest that the fusion of autophagosomes and lysosomes promoted by icarrin enhances autophagy and thus reduces ferroptosis. Therefore, icariin may be a potential candidate for the prevention of ferroptosis of VECs and, thus, for the treatment of cardiovascular diseases.

Side-by-Side OCE-Study of Elasticity and SHG-Characterization of Collagen Fibers in Breast Cancer Tissue before and after Chemotherapy

Side-by-Side OCE-Study of Elasticity and SHG-Characterization of Collagen Fibers in Breast Cancer Tissue before and after Chemotherapy

#6508 THE EFFECTS AND MECHANISM OF BRAIN-TARGETING DRUG-LOADED EXOSOMES ON ADRIAMYCIN NEPHROPATHY MODEL

Abstract Background and Aims Chronic kidney disease (CKD) is a common clinical syndrome, and the current clinical efficacy is still limited. Our previous study confirmed that a small amount of losartan injected into the central lateral ventricle can effectively protect the kidneys, but the route of administration limits its clinical application. As a drug carrier, exosomes have several advantages and are expected to support drug delivery. Therefore, this study aims to develop a brain-targeting exosome complex to provide a new idea for the treatment of CKD. Method In this study, the brain-targeting exosome complex (ExoACP) was synthesized by co-cultivation. The markers of ExoACP were detected by immunoblotting. The structure and particle size distribution were observed by TEM and NTA. Angiotensin II receptor antagonist losartan (ExoACP@Los) was loaded on ExoACP by electroporation. The mice of Adriamycin (ADR)-induced nephropathy were divided into 5 groups: Control, ADR, ADR + Los (IG), ADR + ExoACP (iv.), ADR + ExoACP@Los (iv.). Corresponding treatment was given every other day from the 3rd week after modeling, and all mice were sacrificed at the 5th week, serum and renal tissue was collected for the following experiments. Results This study found that the exosome markers of ExoACP were highly expressed. The Exo and Ang-CP05-Exo were disc-like structures, and the size of the latter was larger than that of the former, suggesting that ExoACP was successfully modified on the surface of Exo. The modification efficiency of ExoACP on Exo was as high as 96.1%. Compared with free Exo, the distribution of ExoACP in brain was increased. Blood biochemical results showed that ExoACP and ExoACP@Los had no apparent side-effects on mice (Figure 1). In-vivo, ExoACP@Los treatment significantly improved the levels of kidney function in mice with ADR nephropathy. ExoACP@Los could significantly reduce renal tubular damage, interstitial fibrosis area and degree of glomerular sclerosis, suggesting that ExoACP@Los could improve the injury of renal tissue (Figure 2). Conclusion This study synthesized a drug-loaded exosome complex with brain targeting and good biocompatibility, and significantly improved renal function, renal tissue damage and collagen fiber deposition in the ADR model.

Visual Observation of Abdominal Adhesion Progression Based on an Optimized Mouse Model of Postoperative Abdominal Adhesions

Background: There is no clear description of the evolution of the progression of abdominal adhesions over time. Method: The optimized model was selected using different adhesion scoring systems. Then, this model was used to observe the progression of abdominal adhesions. Visualized observation of abdominal adhesion evolution was performed by laparoscopy and computed tomography. The inflammatory cell infiltration and collagen fibers in adhesion tissues at different times were evaluated by hematoxylin-eosin and picrosirius red staining. RNA sequencing was used to predict potential key targets of abdominal adhesions at different times. Results: The abdominal adhesion model showed the highest reproducibility when it was established using a circular tool and an electric brush. Based on this model, we found that the inflammatory response was activated early in the process of adhesion formation, peaking on day 3 and then gradually decreasing until stabilization on day 7. Collagen and fibronectin formed on day 1 and gradually increased until remaining stable on day 7. In addition, the characteristic changes in the adhesion zone from initial congestion, edema and fragile tissue to later dense and stable tissue could be vividly observed in live mice by laparoscopy and artificial pneumoperitoneum CT. The RNA sequencing results revealed that Hck on day 1, Ndufs3 and Ndufs8 on day 3 and Aif1 on day 7 might play key roles in abdominal adhesion formation. Conclusion: The construction of a standard process for describing the evolution of abdominal adhesions based on an optimized mouse model will help to facilitate subsequent adhesion-related studies.

Generative adversarial network based digital stain conversion for generating RGB EVG stained image from hyperspectral H&E stained image.

Quantification of elastic fiber in the tissue specimen is an important aspect of diagnosing different diseases. Though hematoxylin and eosin (H&E) staining is a routinely used and less expensive tissue staining technique, elastic and collagen fibers cannot be differentiated using it. So, in conventional pathology, special staining technique, such as Verhoeff's van Gieson (EVG), is applied physically for this purpose. However, the procedure of EVG staining is very expensive and time-consuming. The goal of our study is to propose a deep-learning-based computerized method for the generation of RGB EVG stained tissue from hyperspectral H&E stained one to save the time and cost of conventional EVG staining procedure. H&E stained hyperspectral image and EVG stained RGB whole slide image of human pancreatic tissue have been leveraged for this experiment. CycleGAN-based deep learning model has been proposed for digital stain conversion while images from source and target domains are of different modalities (hyperspectral and RGB) with different channel dimensions. A set of three basis functions have been introduced for calculating one of the losses of the proposed method, which retains the relevant features of EVG stained image within the reduced channel dimension of the H&E stained one. The experimental results showed that a set of three basis functions including linear discriminant function and transmittance spectrum of eosin and hematoxylin better retained the essential properties of the elastic fiber to be discriminated from collagen fiber within the reduced dimension of the hyperspectral H&E stained image. Also, only a smaller number of paired training data with our proposed training method contributed significantly to the generation of more realistic EVG stained image with more precise identification of elastic fiber. RGB EVG stained image is generated from hyperspectral H&E stained image for which our model has performed two types of image conversion simultaneously: hyperspectral to RGB and H&E to EVG. The experimental results show that the intentionally designed set of three basis functions contains more relevant information and prove the effectiveness of our proposed method in generating realistic RGB EVG stained image from hyperspectral H&E stained one.

Establishment of a novel co-cultured liver cancer model based on tumor microenvironment

Objective: To construct a new co-cultured liver cancer research model composed of activated hepatic stellate cells (aHSC) and liver cancer cells, explore the efficacy difference between it and traditional model, so as to establish a liver cancer research model in vitro and in vivo that can reflect the real clinical efficacy. Methods: A new co-culture model of liver cancer consisting of aHSC and liver cancer cells was constructed. The differences in efficacy between the new co-culture model and the traditional single cell model were compared by cytotoxicity test, cell migration test, drug retention test and in vivo tumor inhibition test. Western blot was used to detect the drug-resistant protein P-gp and epithelial-mesenchymal transition-related proteins. Masson staining was used to observe the deposition of collagen fibers in tumor tissues of tumor-bearing mice. CD31 immunohistochemical staining was used to observe the microvessel density in tumor tissues of tumor-bearing mice. Results: The cytotoxicity of single cell model and co-culture model was dose-dependent. With the increase of curcumin (CUR) concentration, the cell viability decreased, but the cell viability of single cell model decreased faster than that of co-culture model. When the concentration of CUR was 10 μg/ml, the cell viability of the co-culture model was 62.3% and the migration rate was (28.05±3.68)%, which were higher than those of the single cell model [38.5% and (14.91±5.92)%, both P<0.05]. Western blot analysis showed that the expressions of P-gp and vimentin were up-regulated in the co-culture model, which were 1.55 and 2.04 fold changes of the single cell model, respectively. The expression of E-cadherin was down-regulated, and the expression level of E-cadherin in the single cell model was 1.17 fold changes of the co-culture model. Drug retention experiment showed that the co-culture model could promote drug efflux and reduce drug retention. In vivo tumor inhibition experiment showed that the m-HSC+ H22 co-transplantation model had faster tumor growth and larger tumor volume than those of the H22 single cell transplantation model. After CUR treatment, the tumor growths of m-HSC+ H22 co-transplantation model and H22 single cell transplantation model were inhibited. Masson staining showed that the deposition of collagen fibers in tumor tissues of m-HSC+ H22 co-transplantation model mice was more than that of H22 single cell transplantation model. CD31 immunohistochemical staining showed that the microvessel density in tumor tissue of m-HSC+ H22 co-transplantation model was higher than that of H22 single cell transplantation model. Conclusions: The aHSC+ liver cancer cell co-culture model has strong proliferation and metastasis ability and is easy to be resistant to drugs. It is a new type of liver cancer treatment research model superior to the traditional single cell model.

The Role of RNA Methylation Modification Related Genes in Prognosis and Immunotherapy of Colorectal Cancer.

Researches showed RNA methylation genes can affect the prognosis of tumors. Thus, the study aimed to comprehensively analyze the effects of RNA methylation regulatory genes in prognosis and treatment of colorectal cancer (CRC). Prognostic signature associated with CRCs were constructed by differential expression analysis, Cox and Least Absolute Shrinkage and Selection Operator (LASSO) analyses. Receiver operating characteristic (ROC) and Kaplan-Meier survival analyses were used to validate the reliability of the developed model. Gene Ontology (GO), Gene set variation analysis (GSVA), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed for functional annotation. Finally, normal and cancerous tissue were collected to validate gene by quantitative real-time PCR (qRT-PCR). A prognostic risk model based on leucine rich pentatricopeptide repeat containing (LRPPRC) and ubiquitin-like with PHD and ring finger domains 2 (UHRF2) was constructed and relevant to the overall survival (OS) of CRC. Functional enrichment analysis revealed that collagen fibrous tissue, ion channel complex and other pathways were significantly enriched, which might help explain the underlying molecular mechanisms. There were significant differences in ImmuneScore, StromalScore, ESTIMATEScore between high- and low-risk groups (p < 0.05). Ultimately, qRT-PCR validation showed that a significant upregulation in the expression of LRPPRC and UHRF2 in cancerous tissue, which verified the effectiveness of our signature. In conclusion, 2 prognostic genes (LRPPRC and UHRF2) related to RNA methylation were identified by bioinformatics analysis, which might supply a new insight into the treatment and evaluation of CRC.

Protective effect of (Andrographis Paniculata) on 4-Vinylcyclohexene Diepoxide Induced ovarian Toxicity in Female Albino Rats

Abstract&#x0D; Background and objective: Protective effect of (Andrographis Paniculata) on 4-Vinylcyclohexene Diepoxide.&#x0D; Material and methods:A group of 55 female albino rats undergo experiment on the outcome effect of 4-vinylcyclohexene Diepoxide injection for a priod of two weeks, as well as treatment effect of methanolic leaf extract of Andrographis paniculata oral administration for four weeks. Injection concentrations (160 and 320mg/kg. B.W) also two different doses of treatment (100 and 200mg/kg. B.W) have been applied.&#x0D; Results:The results on body weight and ovary weight were more evident when higher concentration dose was applied. Whereas, both induced dose were effective and no significant difference was recorded. Histological examination of ovary induced clearly the effect in various ovary tissues, and accumulation of connective tissue collagen fibers around corpus luteum where as a higher dose of treatment found to be behind sign of re-normalizing the within four weeks of treatment.&#x0D; Conclusion: More detailed study proposed to findout the exact right dose for treatment and may the further study extend to include other medicinal plant that might be more efficient.&#x0D; Keywords: 4-Vinylcyclohexene Diepoxide(VCD), Andrographis Paniculata Leaf, Ovarian Toxicity, Rats.

Abstract P5-02-09: Quantitative analysis of fiber-level collagen features in H&amp;E whole-slide images predicts neoadjuvant therapy response in patients with HER2+ breast cancer

Abstract Background: Neoadjuvant treatment (NAT) combining chemotherapy and HER2-targeted agents is frequently administered to HER2-positive (HER2+) breast cancer (BC) patients, with some experiencing a pathological complete response (pCR) and others having residual disease measured by the residual cancer burden (RCB) score. Here, we use a physics-guided machine learning (ML)-based approach to extract fiber-level collagen features from hematoxylin and eosin (H&amp;E)-stained whole slide images (WSIs) and identify collagen-related associations with treatment response in HER2+ patients receiving NAT. Methods: Clinical data and specimens from stage II-III HER2+ BC patients enrolled on the De-escalation to Adjuvant Antibodies Post-pCR to Neoadjuvant THP (DAPHNe; NCT03716180) clinical trial and treated with neoadjuvant paclitaxel/trastuzumab/pertuzumab were analyzed. An ML-based model trained to identify regions of BC tissue as invasive carcinoma, ductal carcinoma in situ (DCIS), diffuse inflammatory infiltrate, stroma, necrosis, or normal tissue was deployed on WSIs of H&amp;E-stained diagnostic core needle biopsies (N=89) to generate tissue overlays. Additional tissue areas were computed from the tissue model predictions using heatmap transformation, including tumor nests (continuous regions predicted as invasive cancer epithelium or DCIS), tumor nest borders (stromal region boundaries 10 μm from tumor nests), and bulk tumor borders (stromal region boundaries 300 μm from aggregated tumor nests). A separate ML-based model trained to identify fiber-level collagen features in WSIs of H&amp;E-stained specimens was also deployed to generate collagen overlays. A fiber feature extraction pipeline was utilized to characterize properties of all identified collagen fibers in the WSI (on the order of hundreds of thousands per slide), including length, width, tortuosity, and angle. These fiber features were then assessed based on their position within the tumor (e.g. relative to the tumor nest border). Combinatorial features (e.g. angle of fibers with respect to tumor boundary) were then explored univariately for associations (N=609) with treatment response. Patients with pCR (RCB=0; N=53) were considered responders, while all other cases (RCBI-III; N=36) were designated non-responders. Due to the small size of the cohort analyzed here, raw p-values are reported. Results: Using estrogen receptor status as a clinical covariate, a logistic regression-based univariate analysis of 609 collagen-associated features revealed six features to strongly associate with pCR (p&amp;lt; 0.05, AUC≥0.75; Table 1). Notable feature themes were identified: 1) fiber tortuosity in tumor nest borders and tumor borders, 2) angle of fibers in tumor border with respect to tumor boundary, and 3) distribution patterns of fiber width in tumor nest borders. The presence of fibers perpendicular to tumor boundary tangents was negatively associated with pCR, as was higher fiber tortuosity and thickness in tumor nest borders. Conclusions: Improved prediction of response to NAT in patients with BC is needed to determine appropriate treatment strategies for each patient. Here, using ML-based models to identify tissue features and collagen fibers, we identify collagen-associated features, measured directly from WSIs of H&amp;E-stained diagnostic BC biopsies, that negatively correlate with pCR. Additional development of this strategy, including the addition of cell identification models and known clinical information, is underway to further refine this novel predictive model. Citation Format: Tan H. Nguyen, Mohammad Mirzadeh, Aaditya Prakash, Emma L. Krause, Jun Zhang, Michael Pyle, Esther R. Ogayo, Harry C. Cramer, Busem Binboga Kurt, Jacqueline Brosnan-Cashman, Michael G. Drage, Stuart Schnitt, Andrew H. Beck, Michael Montalto, Ilan Wapinski, Laura Chambre, Sara Tolaney, Adrienne Waks, Justin Lee, Elizabeth A. Mittendorf. Quantitative analysis of fiber-level collagen features in H&amp;E whole-slide images predicts neoadjuvant therapy response in patients with HER2+ breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-02-09.

Identification of biomarkers and prediction of upstream miRNAs in diabetic nephropathy.

To explore biomarkers of diabetic nephropathy (DN) and predict upstream miRNAs. The data sets GSE142025 and GSE96804 were obtained from Gene Expression Omnibus database. Subsequently, common differentially expressed genes (DEGs) of renal tissue in DN and control group were identified and protein-protein interaction network (PPI) was constructed. Hub genes were screened from in DEGs and made an investigation on functional enrichment and pathway research. Finally, the target gene was selected for further study. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficiency of target gene and predicted its upstream miRNAs. 130 common DEGs were obtained through analysis, and 10 Hub genes were further identified. The function of Hub genes was mainly related to extracellular matrix (ECM), collagen fibrous tissue, transforming growth factor (TGF) -β, advanced glycosylation end product (AGE) -receptor (RAGE) and so on. Research showed that the expression level of Hub genes in DN group was significantly higher than that in control group. (all P<0.05). The target gene matrix metalloproteinase 2 (MMP2) was selected for further study, and it was found to be related to the fibrosis process and the genes regulating fibrosis. Meanwhile, ROC curve analysis showed that MMP2 had a good predictive value for DN. miRNA prediction suggested that miR-106b-5p and miR-93-5p could regulate the expression of MMP2. MMP2 can be used as a biomarker for DN to participate in the pathogenesis of fibrosis, and miR-106b-5p and miR-93-5p may regulate the expression of MMP2 as upstream signals.

IL-11 drives the phenotypic transformation of tracheal epithelial cells and fibroblasts to enhance abnormal repair after tracheal injury

Tracheal stenosis (TS) is a multifactorial and heterogeneous disease that can easily lead to respiratory failure and even death. Interleukin-11 (IL-11) has recently received increased attention as a fibrogenic factor, but its function in TS is uncertain. This study aimed to investigate the role of IL-11 in TS regulation based on clinical samples from patients with TS and a rat model of TS produced by nylon brush scraping. Using lentiviral vectors expressing shRNA (lentivirus-shRNA) targeting the IL-11 receptor (IL-11Rα), we lowered IL-11Rα levels in the rat trachea. Histological and immunostaining methods were used to evaluate the effects of IL-11Rα knockdown on tracheal injury, molecular phenotype, and fibrosis in TS rats. We show that IL-11 was significantly elevated in circulating serum and granulation tissue in patients with TS. In vitro, TGFβ1 dose-dependently stimulated IL-11 secretion from human tracheal epithelial cells (Beas-2b) and primary rat tracheal fibroblasts (PRTF). IL-11 transformed the epithelial cell phenotype to the mesenchymal cell phenotype by activating the β-catenin pathway. Furthermore, IL-11 activated the atypical ERK signaling pathway, stimulated fibroblasts proliferation, and transformed fibroblasts into alpha-smooth muscle actin (α-SMA) positive myofibroblasts. IL-11-neutralizing antibodies (IL-11NAb) or ERK inhibitors (U0126) inhibited IL-11 activity and downregulated fibrotic responses involving TGFβ/SMAD signaling. In vivo, IL-11Rα knockdown rats showed unobstructed tracheal lumen, relatively intact epithelial structure, and significantly reduced granulation tissue proliferation and collagen fiber deposition. Our findings confirm that IL-11 may be a target for future drug prevention and treatment of tracheal stenosis.

Effects of short-term high-concentration exposure to PM2.5 on pulmonary tissue damage and repair ability as well as innate immune events

Short-term heavy air pollution still occurs frequently worldwide, especially during the winter heating period in some developing countries, which is usually accompanied by the temporary explosive growth of PM2.5. The pulmonary damage caused by PM2.5 exposure has been determined, but there have been few studies on the repair ability after the cessation of exposure and the important role of innate immune events. This study established a short-term (30 days) high-concentration (15 mg/kg body weight) PM2.5 exposure and recovery (15 days of exposure cessation) model by intratracheal instillation. The results showed that short-term PM2.5 exposure increased the content of collagen fiber in rat lung tissue, which was significantly repaired after recovery by 15 days of exposure cessation. Meanwhile, exposure to PM2.5 also caused changes in lung epithelial function, macrophage polarization and cell autophagy function. Most of these changes could be restored or reversed to a certain extent after recovery. However, there were also some biomarkers, including CLDN18.1, SP-A, SP-D, iNOS, CD206, Beclin1, p62 and LC3B, that were still significantly different between the exposure and control groups after recovery, suggesting that some toxic effects, especially epithelial function damage, were not completely repaired. In addition, there was a significant correlation between pulmonary fibrosis and innate immunity. The present study demonstrated that short-term high-concentration exposure to PM2.5 could cause temporary lung tissue damage and related innate immune events in rats, and the repair ability existed after the cessation of exposure, but part of the damage that required special attention still persisted.

EVALUATION OF DIAGNOSTIC ALGORITHM BASED ON COLLAGEN ORGANIZATION PARAMETERS FOR BREAST TUMORS.

Tumor tissue samples of 5patients with breast fibroadenomas and 20patients with stage I-II BCa were studied. Collagen was identified histochemically by Mallory method. Photomicrographs of thestudied preparations were obtained using a digital microscopy complex AxioScope A1. Morphometric studies were performed using thesoftware CurveAlign v. 4.0. beta and ImageJ. Thealgorithm for determining thequantitative characteristics and spatial organization of thecollagen matrix in tumor tissue samples has been developed and tested. We showed that collagen fibers in theBCa tissue are characterized by significantly lower values of length (p < 0.001) and width (p < 0.001) as well as higher values of straightness (p < 0.001) and angle (p < 0.05) compared to these in thefibroadenoma tissue. No significant difference was found in thedensity of collagen fibers in thetissue of benign and malignant neoplasms of themammary gland. Thealgorithm allows assessing a wide range of parameters of collagen fibers in tumor tissue, including their spatial orientation and mutual arrangement, parametric characteristics and density of thethree-dimensional fibrillar network.

Possibilities of multiphoton microscopy for the diagnosis of the vulvar lichen sclerosus

Vulvar lichen sclerosus (VLS) is a chronic and recurrent dermatosis of an inflammatory nature with severe focal atrophy of the skin. Connective tissue changes are polymorphic and are still not taken into account in histological diagnostics due to the difficulty of interpreting routine histological methods. In this work, we use multiphoton microscopy (MPM) as a new imaging technique that provides detailed information about the organization of collagen fibers in the dermis based on a non-linear second harmonic generation (SHG) process. To determine the degree of connective tissue damage in lichen sclerosus using standard histological techniques and to reveal the diagnostic capabilities of multiphoton microscopy. We studied 42 biopsies with a histopathological diagnosis of VLS and 10 biopsies of normal vulvar skin. Histological, histochemical and immunohistochemical evaluation was used in comparison with MPM data. Quantitative analysis included the determination of the thickness, length of collagen fibers and the average intensity of the SHG signal. A comprehensive study of the skin showed 4 groups of changes that can be regarded as the degree of the dermis damage: initial, mild, moderate, severe. The affected area at the initial and mild degree has subtle changes, however, it is reliably identified by quantitative analysis of the SHG signal. So, the initial degree is characterized by thin (1.3-1.8 µm) long (56-69 µm) collagen fibers, with a moderate degree, the fibers are thickened (3.4-4.3 µm) and fragmented (22-37 µm). The affected area in moderate and severe cases undergoes homogenization, which is associated with the deposition of extremely thin (0.6-0.9 μm) short (16-28 μm) collagen fibers and the expression of type V collagen. Multiphoton microscopy in the second harmonic generation mode is a reliable method for identifying collagen fibers in tissues. The study made it possible to identify 4 degrees of the dermis damage in vulvar lichen sclerosus.

Study on wound healing effect of low-carbon topical dressings with new green packaging

In order to verify the effect of the new green and low-carbon hydrogel dressing on promoting wound healing, this project applied a sodium alginate hydrogel dressing product containing Escherichia coli and taro toxin analgesic polypeptide (The specific ingredients of the dressing) to skin wounds in common rats. Effects of the hydrogel dressing on promoting skin wound healing was evaluated by observing the occurrence and frequency of behavioral changes in rats, observing wwhistological sections under a high-power microscope, changes in serum cytokine indicators, and Image J analysis of collagen fiber reconstruction ratios in tissue sections. Through comprehensive evaluation, it can be found that hydrogel dressing has analgesic, anti-inflammatory and anti-infection effects on rat wound surface, and acts on promoting wound healing, promoting the formation of new blood vessels in the damaged skin tissue area, promoting the growth of granulation tissue, and promoting the reconstruction of collagen fibers in wound tissue.

線維走行と直交するスリット状損傷部に形成されるマウス膝蓋腱治癒組織の巨視的・微視的力学解析

A lack of the application of appropriate mechanical stimuli to cells within healing tissue in tendon defects has been proposed as a possible mechanism of incomplete defect healing with scar formation. This suggested that different mechanical environment in a tendon defect with a different geometry may result in different healing outcome. Therefore, the present study adopted a slit injury model in a mouse patellar tendon model and performed macroscopic and microscopic mechanical analysis on tissue regenerated in the central slit defect. Tensile strength and tangent modulus of the tissue in the defect at 3 weeks of healing were lower significantly than that of normal patellar tendon, which recovered to a level still lower than the normal level but without statistical significance at 6 weeks of healing. Micromechanical analysis on local strain field revealed that microscopic tensile strain was not different between normal patellar tendon and healing tissue at 6 weeks. However, microscopic transverse and shear strain was significantly lower in healing tissue than normal tendon tissue. The former may indicate that collagen fibers in healing tissue exhibit no crimp morphology, and the latter demonstrates that collagen fibers were not aligned to the long axis of the tendon. Although healing tissue in the slit defect possessed better strength that that in a rectangular defect examined in our previous study, microscopic strain field was similar between two models, possibly due to that healing tissue in both models was still immature structurally at 6 weeks of healing when compared to the normal patellar tendon.

Реактивные изменения соединительнотканных структур в периферических нервах овец при патологии печени

Experimental studies were carried out on sheep using a tetramethane model of hepatosis. Ultrastructural data made it possible to characterize morphofunctional changes in peripheral nerves in sheep, in particular, in myelinated and non-myelinated fibers, neurolemmocytes, and connective tissue structures. The pathological process in the peripheral nervous system initiates a combined pathology of the abdominal organs. This is due to the commonality of their innervation with the dorsal and ventral vagus, celiac, phrenic nerves, and branches of the solar plexus. A decrease in the trophic function of the nerves leads to morphological changes in the pancreas, adrenal glands, kidneys, stomach, and small intestine, which were recorded at the ultrastructural level. As a result, hypoxia, energy and plastic insecurity occur, which leads to destructive processes that involve neurolemmocytes, nerve fibers, neurocytes, connective tissue cells and collagen fibers. Structural and functional changes in sheep, with experimental hepatosis, contribute to quantitative and qualitative changes in the nucleoplasm, cytoplasm, swelling and detachment of the karyolemma. Destruction of organelles depends on the intimate processes of cytolysis. At the same time, the connective tissue in the peripheral nerves suffers significantly, and its functional state largely determines the reactivity of the body. It accumulates various substances. An ultrastructural study of clinically healthy animals shows that in the peripheral nerves, in addition to the nerve components, bundles of collagen fibers form a large volume, where they form a dense tie, passing in different directions. Each of them consists of thinner fibrils, which in turn are bundles of subfibrils. Among the fibers are fibroblasts, which have a different degree of differentiation. Thus, mature fibroblasts have a welldeveloped granular endoplasmic reticulum and numerous ribosomes, with an actively functioning Golgi apparatus, mitochondria, and few microfilaments. In pathology, the qualitative composition of fibroblasts changes, lysis of collagen fibers occurs, and communication with neurolemmocytes is lost. At the same time, fibroblasts strengthen contacts with each other. Such contacts between two, three or more fibroblasts are often seen. Others are located next to vascular endothelial cells and pericytes. Apparently, in pathology, fibroblasts require an informative field for the overall coordination of adaptive processes. On the other hand, it is known that fibroblasts begin to produce IL-1, which affects many processes in the body, in particular, it changes the synthetic functions. This is probably why the connective tissue framework in peripheral nerves changes, and their decay, in turn, stimulates the production of IL-1 by fibroblasts. The latter not only destroys connective tissue elements, but also activates the secretion of collagenase, the processes of proliferation and differentiation of fibroblasts. Fibroblasts activated by IL1 produce PGE2.

Achados clínico-patológicos e imuno-histoquímicos em um caso de angiomatose cutânea bovina em uma novilha Holandesa

ABSTRACT: Bovine cutaneous angiomatosis (BCA) is a rare condition characterized by vasoproliferative skin lesions mainly affecting cattle in the northern hemisphere. An eight-month-old Holstein heifer showed two skin easy-bleeding nodules bulging from the epidermis in the forehead and close to the right prescapular region. Skin lesions were within the dermis and had an irregular surface and a soft heterogeneous reddish parenchyma composed of numerous variable-sized arterioles, veins, and capillaries surrounded by abundant collagenous fibrous tissue. Immunohistochemical assays evidenced endothelial cells lining proliferative vascular structure immunolabeled for CD31 and Von Willebrand factor, and vascular smooth muscle cells immunostained for smooth muscle actin. All clinical, pathological, and immunohistochemical features observed in the Holstein heifer were hallmarks of BCA. Considering its potential for hereditary spread, BCA must be included in the differential diagnosis of easily bleeding skin nodules that do not respond to routine topical wound treatments to prevent its spread in Brazilian herds.

Thymofibrolipoma: a case report and review of the literature

BackgroundThymofibrolipoma has been described as a variant of thymolipoma. To date, 3 cases have been reported, and the lesion have been described to consist of extensive areas of collagenous tissue interspersed with islands of mature adipose tissue and strands of thymic tissue.Case presentationA 43-year-old woman had an anterior mediastinal tumor. Macroscopically, the cut surface of the tumor was composed of a yellowish lipomatous component and a uniform whitish fibrous component with elastic stiffness. Microscopically, the tumor was composed of collagenous fibrous tissue with sparse spindle cells, mature adipocytes and strands or islands of thymic tissue. The spindle cells in the fibrous tissue had monoallelic deletion of the 13q14 region and corresponding loss of RB1 and FOXO1A protein expression.ConclusionsThis case report may strengthen the hypothesis that thymofibrolipoma is a neoplastic lesion and a variant of thymolipoma and that thymofibrolipoma and lipofibroadenoma are different names for the same lesion. The name “lipofibroadenoma” was given to the lesion because of its histological resemblance to fibroadenoma of the mammary gland. However, this name does not reflect the pathogenesis of this lesion, and the name “thymofibrolipoma” would be preferable. It will be necessary to discuss whether lipofibroadenoma should be listed as an independent entity in the WHO classification.