scholarly journals Single-cell Transcriptomic Analysis of Eutopic Endometrium and Ectopic Lesions of Adenomyosis

2020 ◽  
Author(s):  
Zhiyong Liu ◽  
Zhonghua Sun ◽  
Hongyun Liu ◽  
Weipin Niu ◽  
Xin Wang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cell Motility ◽  
Single Cell ◽  
Periodic Acid ◽  
Expression Patterns ◽  
Vasculogenic Mimicry ◽  
Cell Subpopulation ◽  
Periodic Acid Schiff ◽  
Eutopic Endometrium ◽  
Invasion And Migration

Abstract Background: Adenomyosis (AM) is a common benign chronic gynaecological disorder; however, the precise pathogenesis of adenomyosis is still poorly understood. Single-cell RNA sequencing (scRNA-seq) can uncover rare subpopulations, explore genetic and functional heterogeneity, and reveal the uniqueness of each cell. It provides us a new approach to reveal biological issues from a more detailed and microscopic perspective. Here, we utilize this revolutionary technology to identify the changes of gene expression patterns between ectopic lesions and the eutopic endometrium at the single-cell level and explore a potential novel pathogenesis of AM.Methods: A control endometrium (sample with leiomyoma excluding endometrial disorders, n=1), eutopic endometrium and ectopic lesion (from a patient with adenomyosis, n=1) samples were analysed by scRNA-seq, and additional leiomyoma (n=3) and adenomyosis (n=3) samples were used to confirm colocalization and vasculogenic mimicry (VM) formation. Protein colocalization was visualized by immunofluorescence, and CD34-periodic acid-Schiff (PAS) double staining was used to assess the formation of VM.Results: The scRNA-seq results suggest that cancer-, cell motility- and inflammation- (CMI) associated terms, cell proliferation and angiogenesis play important roles in the progression of AM. Moreover, the colocalization of EPCAM and PECAM1 increased significantly in the ectopic endometrium group (P < 0.05), cell subpopulation with high copy number variation (CNV) levels possessing tumour-like features existed in the ectopic lesion sample, and VNN1- and EPCAM-positive cell subcluster displayed active cell motility in endometrial epithelial cells. Furthermore, the epithelial cells transformed to endothelial cells with the obvious accumulation of vasculogenic mimicry formations (positively stained with PAS but not CD34, P < 0.05) in ectopic lesions.Conclusions: In the present study, our results support the theory of adenomyosis derived from the invasion and migration of the endometrium. Moreover, cell subcluster with high CNV level and tumour-associated characteristics is identified. Furthermore, epithelial-endothelial transition (EET) and the formation of VM in tumours, the latter of which facilitates the blood supply and plays an important role in maintaining cell growth, were also confirmed to occur in AM. These results indicated that the inhibition of EET and VM formation may be a potential strategy for AM management.

scholarly journals Single-cell transcriptomic analysis of eutopic endometrium and ectopic lesions of adenomyosis

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Zhiyong Liu ◽  
Zhonghua Sun ◽  
Hongyun Liu ◽  
Weipin Niu ◽  
Xin Wang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cell Motility ◽  
Single Cell ◽  
Periodic Acid ◽  
Expression Patterns ◽  
Vasculogenic Mimicry ◽  
Cell Subpopulation ◽  
Periodic Acid Schiff ◽  
Eutopic Endometrium ◽  
Invasion And Migration

Abstract Background Adenomyosis (AM) is a common benign chronic gynaecological disorder; however, the precise pathogenesis of adenomyosis is still poorly understood. Single-cell RNA sequencing (scRNA-seq) can uncover rare subpopulations, explore genetic and functional heterogeneity, and reveal the uniqueness of each cell. It provides us a new approach to reveal biological issues from a more detailed and microscopic perspective. Here, we utilize this revolutionary technology to identify the changes of gene expression patterns between ectopic lesions and the eutopic endometrium at the single-cell level and explore a potential novel pathogenesis of AM. Methods A control endometrium (sample with leiomyoma excluding endometrial disorders, n = 1), eutopic endometrium and ectopic lesion (from a patient with adenomyosis, n = 1) samples were analysed by scRNA-seq, and additional leiomyoma (n = 3) and adenomyosis (n = 3) samples were used to confirm colocalization and vasculogenic mimicry (VM) formation. Protein colocalization was visualized by immunofluorescence, and CD34-periodic acid-Schiff (PAS) double staining was used to assess the formation of VM. Results The scRNA-seq results suggest that cancer-, cell motility- and inflammation- (CMI) associated terms, cell proliferation and angiogenesis play important roles in the progression of AM. Moreover, the colocalization of EPCAM and PECAM1 increased significantly in the ectopic endometrium group (P < 0.05), cell subpopulation with high copy number variation (CNV) levels possessing tumour-like features existed in the ectopic lesion sample, and VNN1- and EPCAM-positive cell subcluster displayed active cell motility in endometrial epithelial cells. Furthermore, during the transformation of epithelial cells to endothelial cells, we observed the significant accumulation of VM formation (positively stained with PAS but not CD34, P < 0.05) in ectopic lesions. Conclusions In the present study, our results support the theory of adenomyosis derived from the invasion and migration of the endometrium. Moreover, cell subcluster with high CNV level and tumour-associated characteristics is identified. Furthermore, epithelial-endothelial transition (EET) and the formation of VM in tumours, the latter of which facilitates the blood supply and plays an important role in maintaining cell growth, were also confirmed to occur in AM. These results indicated that the inhibition of EET and VM formation may be a potential strategy for AM management.

scholarly journals Mucinous Degeneration of the Epithelium of the Urinary Tract of Swine

1971 ◽  
Vol 8 (5-6) ◽  
pp. 485-489 ◽  
Author(s):  
D. F. Brobst ◽  
R. Cottrell ◽  
A. Delez
Keyword(s):  
Urinary Tract ◽  
Epithelial Cells ◽  
Urinary Bladder ◽  
Periodic Acid ◽  
Degenerative Change ◽  
Colloidal Iron ◽  
Periodic Acid Schiff ◽  
Hog Cholera ◽  
Transitional Cells ◽  
Suppurative Arthritis

Mucinous degenerative change was observed in the epithelial cells lining the renal pelvis, ureter, and urinary bladder of pigs with exudative epidermitis, coliform enteritis, hog cholera, and suppurative arthritis. Mucins were observed within transitional cells either as granular or homogenous material within vacuoles. Lakes filled with mucins also were formed as a result of the coalescence of mucin from degenerating transitional cells. The cells and lakes of mucin were stained selectively by periodic acid-Schiff, alcian blue, and colloidal iron. On the basis of the reactivity patterns with these stains the transitional epithelial cells were considered capable of producing acidic and neutral mucins.

scholarly journals GAMBARAN MIKROSKOPIK GINJAL TIKUS WISTAR (RATTUS NORVEGICUS) SETELAH DIINDUKSI DENGAN GENTAMISIN

2013 ◽  
Vol 4 (3) ◽  
Author(s):  
Poppy M Lintong ◽  
Carla F Kairupan ◽  
Priska L N Sondakh
Keyword(s):  
Body Weight ◽  
Epithelial Cells ◽  
Wistar Rats ◽  
Periodic Acid ◽  
Basal Membrane ◽  
Tubular Epithelial Cells ◽  
Periodic Acid Schiff ◽  
Group Iii ◽  
Group I ◽  
Group Ii

Abstract: Gentamycin, a frequently used aminoglycoside antibiotics, has a nephrotoxic effect to human beings and animals. The purpose of this research was to find out the microscopic changes of wistar rat kidneys after gentamycin induction. This was an experimental study, using five adult wistar rats, divided into three groups. Group I was the control group; group II consisted of two rats, injected with gentamycin 0,3 ml/day (dose of 60 mg/kg body weight/day) intraperitoneally for seven days; and group III consisted of two rats, injected with gentamycin 0,3 ml/day intraperitoneally for 10 days. Group I and II were terminated at day-8, and group III at day-11. Their kidneys were processed for microscopic slides, stained with hematoxylin eosin and Periodic Acid Schiff. In microscopic evaluation, group II and III showed oedema, necrosis, apoptosis, and basal membrane destruction of tubular epithelial cells. Group III also showed fat vacuoles in these epithelial cells (macrovesicular fatty changes). Conclusion: wistar rats injected with gentamycin 60 mg/kg body weight/day for 7 and 10 days showed oedema, necrosis, apoptosis, and basal membrane destruction of tubular epithelial cells; and macrovesicular fatty changes after 10 days of gentamycin.Key words: gentamycin, necrosis tubular epithelial cells, fatty changesAbstrak: Gentamisin termasuk antibiotik golongan aminoglikosida berspektrum luas yang bersifat nefrotoksik terhadap manusia dan hewan. Tujuan penelitian ini untuk melihat perubahan mikroskopik struktur ginjal tikus Wistar setelah diberikan gentamisin. Metode penelitian eksperimental dengan menggunakan lima ekor tikus Wistar dewasa yang dibagi atas tiga kelompok. Kelompok I tanpa perlakuan; kelompok II terdiri dari dua ekor tikus perlakuan yang diinjeksi dengan gentamisin 0,3 ml/hari (dosis 60 mg/kgBB/hari) secara intraperitonial selama tujuh hari; dan kelompok III terdiri dari dua ekor tikus perlakuan yang diinjeksi dengan gentamisin 0,3 ml/hari secara intraperitonial selama 10 hari. Tikus Wistar kelompok I dan II diteminasi hari ke-8, sedangkan kelompok III diterminasi hari ke-11. Ginjal tikus kelompok I -III kemudian dibuat preparat histopatologik dengan pengecatan rutin hematoksilin eosin dan Periodic Acid Schiff (PAS). Hasil penelitian menunjukkan tikus Wistar perlakuan yang diberikan gentamisin 0,3 ml/hari selama 7 sampai 10 hari secara mikroskopik memperlihatkan pembengkakan, nekrosis, apoptosis, dan destruksi membrana basalis sel epitel tubulus; dan pada hari ke-10 terlihat vakuol-vakuol lemak pada sel epitel sehingga inti terdesak ke tepi (perlemakan makrovesikuler). Simpulan: pemberian gentamisin pada tikus Wistar dengan dosis 60 mg/kg BB/hari selama 7-10 hari menunjukkan pembengkakan, nekrosis, apoptosis sel epitel tubulus, dan membrana basalis tubulus rusak; dan setelah hari ke-10 juga terlihat perlemakan makrovesikuler.Kata kunci: gentamisin, nekrosis sel epitel tubulus, perlemakan makrovesikuler

scholarly journals Vasculogenic Mimicry Correlates to Presenting Symptoms and Mortality in Uveal Melanoma

2021 ◽  
Author(s):  
Shiva Sabazade ◽  
Viktor Torgny Gill ◽  
Christina Herrspiegel ◽  
Gustav Stålhammar
Keyword(s):  
Visual Field ◽  
Uveal Melanoma ◽  
Poor Prognosis ◽  
Periodic Acid ◽  
Vasculogenic Mimicry ◽  
Clinicopathological Parameters ◽  
Tumor Diameter ◽  
Blurred Vision ◽  
Periodic Acid Schiff ◽  
Presenting Symptoms

Abstract PurposeFluid-conducting extracellular matrix patterns known as vasculogenic mimicry (VM) have been associated with poor prognosis in uveal melanoma and other cancers. We investigate the correlations between VM, presenting symptoms, mortality and the area density of periodic acid-Schiff positive histological patterns (PAS density).MethodsSixty-nine patients that underwent enucleation for uveal melanoma between 2000 and 2007 were included. Clinicopathological parameters, presenting symptoms and outcomes were collected. Histological tumor sections were evaluated for VM and PAS density was quantified with digital image analysis.ResultsThirty-four patients (49 %) presented with blurred vision. 18 (26 %) with a shadow in the visual field, 7 (10 %) with photopsia and/or floaters and 2 (3 %) with metamorphopsia. Nine patients (13 %) had no symptoms at all. Median follow-up was 16.7 years (SD 2.6). A shadow in the visual field, but no other symptom, was positively correlated with the presence of VM (φ 0.70, p<0.001) and greater PAS density (p<0.001). In multivariate regression, retinal detachment (RD), presence of VM and PAS density ≥ median were independent predictors of a shadow, but not tumor distance to the macula, tumor apical thickness, tumor diameter or ciliary body engagement. Presence of VM was associated with significantly shorter cumulative disease-specific survival (Wilcoxon p=0.04), but not PAS density ≥ median, presenting symptoms or RD (p>0.28).ConclusionTumors from uveal melanoma patients that report a visual field shadow are likely to display VM and greater PAS density, likely explaining the previously reported association between this symptom and poor prognosis.

scholarly journals  Histological and histochemical studies on the Harderian gland in native chickens

2012 ◽  
Vol 57 (No. 8) ◽  
pp. 404-409 ◽  
Author(s):  
B. Mobini
Keyword(s):  
Epithelial Cells ◽  
Plasma Cells ◽  
Periodic Acid ◽  
Harderian Gland ◽  
Histochemical Staining ◽  
Alveolar Type ◽  
Periodic Acid Schiff ◽  
Parasympathetic Ganglia ◽  
Tissue Sections ◽  
Native Chickens

&nbsp; The objective of this investigation was to study the histological and histochemical structure of the Harderian gland in native chickens. Samples were obtained from 10 male and 10 female adult healthy native chickens. Tissue sections were stained with haematoxylin eosin, Verhoeff&rsquo;s, Masson&rsquo;s trichrome, alcian blue (pH&nbsp;2.5), periodic acid-Schiff and Gomori&rsquo;s method for reticulum. The multilobular Harderian gland of native chickens was covered by a thin connective tissue which consisted of adipose tissue, parasympathetic ganglia, nerve bundles, collagen, elastic and reticular fibres. Plasma cells were present in interlobular areas. The Harderian gland was compound tubulo-alveolar type. The Harderian duct was lined by columnar epithelial cells of varying height. Goblet cells were not found in Harderian duct. Histochemical staining revealed that the all epithelial cells of both corpus glandulae and ducts contained both neutral and acidic mucins. No significant sex-based differences were found. It is concluded that the general histological and histochemical structure of the Harderian gland in native chickens is similar to that of domestic geese, but that there are also some differences. &nbsp;

scholarly journals Multiple Hepatic Peribiliary Cysts in a Young Pig

2007 ◽  
Vol 44 (5) ◽  
pp. 707-709 ◽  
Author(s):  
M. Komine ◽  
K. Kawasako ◽  
Y. Akihara ◽  
Y. Shimoyama ◽  
M. Okamoto ◽  
...  
Keyword(s):  
Bile Duct ◽  
Epithelial Cells ◽  
Alcian Blue ◽  
Periodic Acid ◽  
Single Layer ◽  
Bile Pigment ◽  
Periodic Acid Schiff ◽  
Neutral Mucin ◽  
Peribiliary Cysts ◽  
Large Bile Duct

Histopathologic features of hepatic peribiliary cysts were described in a young slaughtered pig. The animal was an apparently healthy 6–month-old pig of mixed breed. Macroscopically, all lobes of the liver contained numerous cysts of varying size containing serous fluid in all lobes. Histopathologically, the cysts were located mainly around the large bile duct and in the connective tissue of the portal tracts. Within serial sections, these cysts were assumed to be solitary or multilocular, but they were separated from the bile duct. The cysts were lined by a single layer of columnar, cuboidal, and flattened epithelial cells. Occasionally, goblet cells were observed. The epithelial cells were stained with periodic acid—Schiff/alcian blue and high-iron diamine/alcian blue, indicating the presence of neutral mucin, sialomucin, and sulfomucin. Grimalius' method revealed the presence of endocrine cells in the lining epithelium. There was no bile pigment in the cysts by the Hall method.

scholarly journals Identification of a basal stem cell subpopulation in the prostate via functional, lineage tracing and single-cell RNA-seq analyses

2019 ◽  
Author(s):  
Xue Wang ◽  
Haibo Xu ◽  
Chaping Cheng ◽  
Zhongzhong Ji ◽  
Huifang Zhao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Epithelial Cells ◽  
Single Cell ◽  
Basal Cell ◽  
Lineage Tracing ◽  
Cell Subpopulation ◽  
Basal Cells ◽  
Mesenchymal Transition ◽  
Genetic Ablation

AbstractThe basal cell compartment in many epithelial tissues such as the prostate, bladder, and mammary gland are generally believed to serve as an important pool of stem cells. However, basal cells are heterogenous and the stem cell subpopulation within basal cells is not well elucidated. Here we uncover that the core epithelial-to-mesenchymal transition (EMT) inducer Zeb is exclusively expressed in a prostate basal cell subpopulation based on both immunocytochemical and cell lineage tracing analysis. The Zeb1+prostate epithelial cells are multipotent prostate basal stem cells (PBSCs) that can self-renew and generate functional prostatic glandular structures with all three epithelial cell types at the single-cell level. Genetic ablation studies reveal an indispensable role for Zeb1 in prostate basal cell development. Utilizing unbiased single cell transcriptomic analysis of over 9000 mouse prostate basal cells, we find that Zeb1+basal cell subset shares gene expression signatures with both epithelial and mesenchymal cells and stands out uniquely among all the basal cell clusters. Moreover, Zeb1+epithelial cells can be detected in mouse and clinical samples of prostate tumors. Identification of the PBSC and its transcriptome profile is crucial to advance our understanding of prostate development and tumorigenesis.

scholarly journals Combined transient ablation and single cell RNA sequencing reveals the development of medullary thymic epithelial cells

2020 ◽  
Author(s):  
Kristen L. Wells ◽  
Corey N. Miller ◽  
Andreas R. Gschwind ◽  
Wu Wei ◽  
Jonah D. Phipps ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Single Cell ◽  
Rna Sequencing ◽  
Expression Patterns ◽  
Critical Role ◽  
Lineage Tracing ◽  
Thymic Epithelial Cells ◽  
Single Cell Rna Sequencing ◽  
Medullary Thymic Epithelial Cells

AbstractMedullary thymic epithelial cells (mTECs) play a critical role in central immune tolerance by mediating negative selection of autoreactive T cells through the collective expression of the peripheral self-antigen compartment, including tissue-specific antigens (TSAs). Recent work has shown that gene expression patterns within the mTEC compartment are remarkably heterogenous and include multiple differentiated cell states. To further define mTEC development and medullary epithelial lineage relationships, we combined lineage tracing and recovery from transient in vivo mTEC ablation with single cell RNA-sequencing. The combination of bioinformatic and experimental approaches revealed a non-stem transit-amplifying population of cycling mTECs that preceded Aire expression. Based on our findings, we propose a branching model of mTEC development wherein a heterogeneous pool of transit-amplifying cells gives rise to Aire- and Ccl21a-expressing mTEC subsets. We further use experimental techniques to show that within the Aire-expressing developmental branch, TSA expression peaked as Aire expression decreased, implying Aire expression must be established before TSA expression can occur. Collectively, these data provide a higher order roadmap of mTEC development and demonstrate the power of combinatorial approaches leveraging both in vivo models and high-dimensional datasets.

scholarly journals Single-Cell Transcriptome Analysis of Human Adipose-Derived Stromal Cells Identifies a Contractile Cell Subpopulation

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Elize Wolmarans ◽  
Juanita Mellet ◽  
Chrisna Durandt ◽  
Fourie Joubert ◽  
Michael S. Pepper
Keyword(s):  
Single Cell ◽  
Transcriptome Analysis ◽  
Stromal Cells ◽  
Exploratory Study ◽  
Expression Patterns ◽  
Cell Subpopulation ◽  
Adipose Derived Stromal Cells ◽  
Cell Transcriptome ◽  
Population Doublings ◽  
Single Cell Transcriptome

The potential for human adipose-derived stromal cells (hASCs) to be used as a therapeutic product is being assessed in multiple clinical trials. However, much is still to be learned about these cells before they can be used with confidence in the clinical setting. An inherent characteristic of hASCs that is not well understood is their heterogeneity. The aim of this exploratory study was to characterize the heterogeneity of freshly isolated hASCs after two population doublings (P2) using single-cell transcriptome analysis. A minimum of two subpopulations were identified at P2. A major subpopulation was identified as contractile cells which, based on gene expression patterns, are likely to be pericytes and/or vascular smooth muscle cells (vSMCs).

Submucosal Lifting Agent ORISE Gel Remnants Histopathologically Mimic Mucin and Malignancy: A Case Series

2019 ◽  
Vol 152 (Supplement_1) ◽  
pp. S73-S73 ◽  
Author(s):  
Luke Cypher ◽  
Shaoli Sun ◽  
Erin Forster ◽  
Brenda Hoffman ◽  
David Lewin
Keyword(s):  
Colon Cancer ◽  
Epithelial Cells ◽  
Amorphous Material ◽  
Periodic Acid ◽  
Case Series ◽  
The United States ◽  
Periodic Acid Schiff ◽  
Good Communication ◽  
Gastrointestinal Lesions ◽  
Hematoxylin And Eosin

Abstract Introduction There were an estimated 18.1 million new cancer cases in 2018, with colon cancer being the third most common worldwide. Colon cancer development is an accumulation of mutations resulting in normal epithelial cells transforming into adenomas and then adenocarcinomas. In certain scenarios, endoscopic interventions have gained considerable momentum over invasive surgery as an alternative to manage early gastrointestinal lesions. New techniques such as endoscopic mucosal resection (EMR) and endoscopic submucosal dissection allow for removal of large, flat sessile polyps. Successful EMR is dependent on expanding the submucosal space to create adequate lift of the polyp to facilitate tissue capture and to avoid perforation and excess bleeding. ORISE gel (Boston Scientific) is a submucosal lifting agent currently in use in the United States. Methods We present three cases of gastrointestinal specimens obtained using ORISE gel. Histological analysis with hematoxylin and eosin revealed submucosal amorphous deposits that appeared to be mucin. Due to the concern for malignancy, additional stains were performed, including periodic acid–Schiff with diastase digestion (DPAS) to identify mucin. DPAS staining for mucin was negative, indicating the mucinous-appearing amorphous material seen on hematoxylin and eosin staining was not mucin but a likely remnant ORISE gel used during EMR. Additional immunohistochemical stains for epithelial cells (cytokeratin AE1/AE3) were also performed to exclude the presence of infiltrating tumor cells. Conclusion These three cautionary cases reveal the importance of good communication between endoscopists and pathology. In an effort to avoid overdiagnosis and/or the usage of unnecessary additional stains, pathologists should be alerted of ORISE gel usage.

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