Number Of Epithelial Cells Research Articles

Abstract 3045: Role of CD10 negative stroma in the development of precursor lesion of endometriosis associated ovarian cancer

Abstract Introduction: Ovarian clear cell carcinoma (OCCC) is a rare, aggressive, chemo-resistant subtype of epithelial ovarian cancer, and is associated with a significantly worse five-year survival compared to other ovarian cancer types. This is in part due to its distinct cell of origin which is thought to arise within sites of endometriosis. Atypical endometriosis is considered a precursor lesion of endometriosis-associated ovarian cancers, including OCCC. However, the mechanism underlying the development of atypical endometriosis and subsequent malignant transformation is still unknown. We recently demonstrated the existence of a OCCC tumor-promoting mesenchymal stem cell (enMSCs), marked by the loss of CD10 expression, within the endometriotic microenvironment. Further, we demonstrated that CD10 negative enMSCs selectively enhance OCCC growth, but not high-grade serous ovarian cancer (HGSOC) growth which provides further evidence that the endometriotic microenvironment is uniquely supportive of OCCC tumorigenesis. Thus, we hypothesized that loss of stromal CD10 expression leads to the creation of the precursor microenvironment for OCCC. Methods: EnMSCs and endometriosis epithelial cells were isolated from endometriosis tissue. Tissues were collected from women who undergo surgery for benign endometriosis. Based on surface CD10 expression, enMSCs were divided into CD10 negative or positive populations. Endometriosis epithelium proliferation was assessed using in vitro assays. Long term organoid of CD10 negative enMSCs and endometriosis epithelium were implanted into NSG mice to assess tumor initiation capacity. Results: Our data showed that co-culture with CD10 negative enMSCs enhanced the proliferation of endometriosis epithelium. Further, CD10 negative enMSCs enhanced the sphere formation capacity (sphere number and sphere size) of endometriosis epithelial cells when compared to endometriosis epithelium co-cultured with CD10 positive enMSCs or cultured alone. Furthermore, long term growth of organoids containing CD10 negative enMSCs and endometriosis epithelium stimulated the development of cytologic atypia in the epithelial cells. Strikingly, these organoids, when injected into the fat pad of NSG mice, initiated localized tumors. In summary, our data highlighted the role of CD10 negative enMSCs in the development of cytologic atypia in endometriosis epithelium and inducing tumor formation in vivo. This work has the potential for significant clinical impact as identification of a stromal biomarker for OCCC development would have important implications in the clinical management of women with endometriosis, including determining the need for risk-reducing surgery. Further, this work has the potential to identify new targets to disrupt the unique microenvironment of OCCC, thus providing new therapeutic strategies for this deadly disease. Citation Format: Huda Issa Atiya, Lan Coffman. Role of CD10 negative stroma in the development of precursor lesion of endometriosis associated ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3045.

Assessment of Cytological Changes in the Oral Mucosa in Young Hematological Patients Treated with Systemic Chemotherapy

The primary objective of the undertaken study was to determine the morphological changes that occur within the oral epithelium in children undergoing chemotherapy following a diagnosis of hematological malignancies. The study group consisted of 18 patients diagnosed with leukemia or lymphoma undergoing treatment with chemotherapy. Swabs (liquid-based cytology) were collected from the oral cavity for microscopic evaluation at baseline, during the chemotherapy cycle with oral mucositis symptoms present, and upon completion of the cycle. Both the neutrophil count and oral mucositis (OM) were registered using the WHO (World Health Organization) scale. The control group included 41 children who were generally healthy. All samples underwent microscopical analyses at the Department of Pathology, Medical University of Gdansk, Poland. A total of 190 cytological preparations were evaluated. The baseline preparations revealed similar cytological images, and the superficial cells of the epithelial layers were seen. A significant (p < 0.01) increase in the number of cells in the intermediate layer of the oral epithelium, as well as a decrease (p < 0.01) in the volume of cells in the superficial layers, was observed in further stages of cytostatic treatment. A decrease in the percentage of superficial epithelial cells with a corresponding increase in the number of intermediate epithelial cells is considered to be a result of toxic damage to the oral mucosa during chemotherapy.

A STUDY ON TRANS URETHRAL RESECTION OF PROSTATE FOR SYMPTOMATIC BPH PRESENTING IN A TERTIARY CARE HOSPITAL

Introduction: Benign prostatic hyperplasia (BPH) is a common chronic progressive disease resulting in urinary obstruction in aging men. It is a pathologic process that consists of androgen-dependent increases in the number of epithelial and stromal cells in the periurethral area of the prostate. BPH contributes to, but is not the sole cause of, lower urinary tract symptoms in aging men. Many patients require surgical treatment and presently TURP (Trans Urethral Resection of Prostate) is considered the gold standard in surgical management of BPH. Aim: To observe and assess the outcome of TURP being done in Narayan Medical College and Hospital Sasaram, Bihar. Materials and Methods: The records of patients who presented to the surgical OPD with LUTS and were assessed to undergo TURP for prostatic hyperplasia. We present our experience with series of 47 patients and can say that TURP can be used for most patients presenting with BPH in peripheral areas. Result: In the study period 47 TURP procedures were performed for BPH. The mean age of the cohort was 67.9 years;( ranging from 56 to 84 years). The mean prostatic volume was 44.06 grams (range 32 to 76 grams) and mean operating time was 48 minutes (range 29 to 85 minutes). Blood transfusion was required in 3 patients (6.3%). No patients required open intervention and there were no mortalities. The common complications were bleeding (12%), UTI and clot retention. Conclusions: TURPis not available to majority of rural population having symptomatic benign prostatic hyperplasia (BPH) due to lack of facility but remains the treatment of choice provided trained surgeons and facilities are there. The overall cost of TURP is marginally higher for an average rural patient as compared to open surgery but is worthwhile in view of its inherent minimal trauma, short hospital stays and early recovery and huge cost advantage over newer therapies like HoLEPor PVP

Prostate-specific antigen density as the best predictor of low- to intermediate-risk prostate cancer: a cohort study.

Evaluation of prostate cancer (PCa) when serum prostate-specific antigen (PSA) level is vaguely elevated is complicated. This is because serum PSA levels only reflect the number of prostate epithelial cells. We aimed to compare PSA and various prostate volume-related factors to determine which one can best predict PCa in patients with a PSA level of 2.5-20 ng/mL. Patients who underwent transrectal ultrasound (TRUS)-guided prostate biopsy at the Inje University Sanggye Paik Hospital between January 2018 and July 2021 and who had a PSA level of 2.5-20 ng/mL were retrospectively identified (n=275). Among them, based on biopsy results, patients were divided into cancer group and non-cancer groups, and age, PSA, total prostate volume (TPV), peripheral zone volume (PZV), peripheral zone PSA density (PZ-PSAD), transitional zone-PSAD (TZ-PSAD), and PSAD were compared and analyzed using receiver operating characteristic (ROC) and univariate analyses. The areas under ROC curves (AUCs) for age, total PSA, TPV, PZV, PZ-PSAD, TZ-PSAD, and PSAD for predicting PCa in patients with a PSA level of 2.5-20.0 ng/mL were 0.678, 0.680, 0.671, 0.639, 0.731, 0.736, and 0.764, respectively. In univariate and multivariate analysis, all categorical variables were divided based on the cut-off value and used to predict PCa. Those with a PSAD of ≥0.218 ng/mL2 were found to be at an increased risk of PCa than those with a PSAD of <0.218 ng/mL2 [odds ratio (OR) =3.51; 95% confidence interval (CI): 1.306-9.415], which was the best result, followed by TZ-PSAD with a cut-off value of 0.353. At a PSAD level of 0.218 ng/mL2, 85.0% of the PCa group could avoid unnecessary biopsy and 61.4% of the non-PCa group could reduce missed diagnosis when the TRUS findings were inaccurate. PSAD may inform biopsy decisions as the best predictor of PCa when TRUS findings are ambiguous in patients with a PSA level of 2.5-20.0 ng/mL.

Establishment of Epithelial Cell Culture from Ovarian Cancer Tissues: A Method Comparison Study.

This study aimed to determine the most effective method in establishing primary cell culture from epithelial serous ovarian cancer tissues with the highest yield of cells and percentage of epithelial cells. Primary and metastasis tissues from three serous ovarian cancer patients were processed using 18 different combinations of methods based on different factors: the source of tissue (primary site or metastasis site), the cell suspension method (explant method, enzymatic methods, or the addition of Percoll), and the alternatives of three different media. We compared the total count of cells, the percentage of epithelial cells, and the estimated number of epithelial cells per observation field. The calculation of cells from primary tissues were compared to metastasis tissues, and the difference was statistically analyzed using Mann Whitney-U test on SPSS software. The groups that were processed using dispase and trypsin resulted higher number of cells and higher percentage of epithelial cells when compared to the explant method. Among all media, we found that DMEM:F12 and McCoy's 5A media as equally useful in isolating and culturing epithelial cells. Statistically, the metastasis tissue derived more epithelial cells when compared to the primary tissue (102.32±82.65 vs 22.6±23.81, p=0.001). The use of metastasis tissue processed with trypsin or dispase and cultured in DMEM:F12 or McCoy's 5A media was found to be the most efficient way to produce the highest amount of cells with high percentage of epithelial cells.

Сучасні відомості про патогенетичні механізми формування пневмосклерозу

The purpose of the study was to analyze literary sources on the study of modern views on information about the pathogenetic mechanisms of the formation of pulmonary fibrosis. Materials and methods. Analytical and bibliosemantic methods were used in the research. During the scientific search, 39 sources of modern domestic and foreign literature were reviewed and analyzed. Results and discussion. Pulmonary fibrosis is a heterogeneous group of chronic, progressive and incurable interstitial lung diseases characterized by scar formation and irreversible destruction of the lung parenchyma and is accompanied by disorders of elasticity and gas exchange in pathologically altered areas. The mechanism of development of pulmonary fibrosis is determined by its root causes. There are three distinct pathologic patterns of pulmonary fibrosis: usual interstitial pneumonia, fibrotic nonspecific interstitial pneumonia, and airway fibrosis. Their morphological differences are based on the distribution of fibrosis (diffuse or spotty) and anatomical location. The development of pulmonary fibrosis in most cases is a consequence of a previous acute inflammation of the lungs caused by various etiological factors, which in the case of untimely started or incorrectly selected treatment causes the deposition of fibrous tissue in the lungs. It is believed that the appearance and subsequent progression of pulmonary fibrosis can be attributed to reparative processes after repeated injuries of alveolar epithelial cells in response to various stimuli, including injuries. Loss of function or reduction in the number of alveolar epithelial cells can lead to improper repair of the lung parenchyma, which can lead to fibrosis. Various cytokines such as transforming growth factor-β1, tumor necrosis factor-α, and platelet-derived growth factor can be released when alveolar epithelial cells are damaged. These cytokines can promote the accumulation of fibroblasts. In addition to the cytokine response, the lung’s response to injury includes the stimulation of myofibroblasts, which when activated serve as the primary collagen-producing cell. This leads to massive deposition of collagen and subsequently affects the normal structure and function of lung tissue. Conclusion. Pulmonary fibrosis is a progressive lung disease that leads to morpho-functional restructuring of lung tissue. In the course of the work, the presence of three models of the development of pulmonary fibrosis were analyzed. Despite the long history of study and good coverage of the problem in the scientific literature, currently the mechanisms of formation of pulmonary fibrosis remain insufficiently studied

Modeling of Respiratory Diseases Evolving with Fibrosis from Organoids Derived from Human Pluripotent Stem Cells

Respiratory disease is one of the leading causes of morbidity and mortality worldwide. There is no cure for most diseases, which are treated symptomatically. Hence, new strategies are required to deepen the understanding of the disease and development of therapeutic strategies. The advent of stem cell and organoid technology has enabled the development of human pluripotent stem cell lines and adequate differentiation protocols for developing both airways and lung organoids in different formats. These novel human-pluripotent-stem-cell-derived organoids have enabled relatively accurate disease modeling. Idiopathic pulmonary fibrosis is a fatal and debilitating disease that exhibits prototypical fibrotic features that may be, to some extent, extrapolated to other conditions. Thus, respiratory diseases such as cystic fibrosis, chronic obstructive pulmonary disease, or the one caused by SARS-CoV-2 may reflect some fibrotic aspects reminiscent of those present in idiopathic pulmonary fibrosis. Modeling of fibrosis of the airways and the lung is a real challenge due to the large number of epithelial cells involved and interaction with other cell types of mesenchymal origin. This review will focus on the status of respiratory disease modeling from human-pluripotent-stem-cell-derived organoids, which are being used to model several representative respiratory diseases, such as idiopathic pulmonary fibrosis, cystic fibrosis, chronic obstructive pulmonary disease, and COVID-19.

Changes in Lower Urinary Tract Symptoms in Benign Prostatic Hyperplasia Patients Affected by Covid-19

Introduction: Benign prostatic hyperplasia (BPH) is a histological diagnosis characterized by an increase in the number of epithelial and stromal cells in the transitional zone of the prostate gland. The clinical manifestation of BPH is associated with the appearance of so-called lower urinary tract symptoms (LUTS) which can also be a consequence of other conditions not related to the prostate. Covid-19, also known as severe acute respiratory syndrome coronavirus 2 was discovered as a disease in late 2019 in the city of Wuhan, China. Materials and Methods: A case-control study was conducted between September 2021 and May 2022. Information was initially collected on 60 patients followed up and treated conservatively for BPH in two outpatient practices and recovered from Covid-19. After processing the received information, 27 patients were excluded from the study and 33 patients remained for observation The assessment of patients is carried out through an internationally validated questionnaire – international prostate symptom score (IPPS), prostate specific antigen, digital rectal examination (DRЕ), and ultrasound diagnostics of the prostate gland with consideration of its volume and the amount of residual urine. Results: Our data show a strong correlation between the changes in LUTS in patients with BPH and Covid-19 infection. Conclusion: Although almost 3 years have passed since the beginning of this pandemic, there are still many unanswered questions surrounding this disease. We believe that with our experience we will enrich the information about the relationship between Covid-19 and LUTS, and the results obtained by us can serve as a basis for future large-scale and more in-depth studies on the subject.

Cytological characteristics of purulent wounds in the oral cavity using antiseptic dressing “Vita Valis”

Aim. To study the effect of antiseptic sorption dressing “Vita Wallis” on the course of the wound process in acute purulent inflammatory diseases of the oral cavity.Materials and methods. We have conducted a cytological examination of 30 patients with the acute odontogenic purulent inflammatory process of the oral cavity. Among them were 17 men (56,7%) and 13 women (43,3%). The patients were divided into two clinical groups equally. Cytological research was carried out on the specimen preparations, which were taken by putting slides to the purulent wounds on the 1st, third and fifth days of the postoperative period.Results and discussion. The cytological study showed that the wound process had a distinct phase character on the first day after the operation in both groups. On the third and fifth day decrease of neutrophils, number was observed in the wounds (control group - from 65,0 to 90,5% and 56,0 to 66,0%, the main group - 52,0 to 80,5% and 40,0 to 53,0%). During this period, the number of epithelial cells in the main group patients increased more than threefold compared with the control group; it varied from 25.0 to 33.5%.Conclusion. Application of antiseptic sorption material “Vita Vallis”, in comparison with the conventional method of treatment, cleanses purulent wounds more intensively, contributing to the intensified migration of inflammatory infiltrate cells and further regeneration in the damaged area.

Open hepatic artery flow with portal vein clamping protects against bile duct injury compared to pringles maneuver

Background Conventional hepatic artery and portal vein clamping strategies can prevent blood loss and ischemia-reperfusion liver injury, and such preventative measures are the key to successful liver surgery. However, ischemic-induced damage to cholangiocytes is rarely considered. Here, we aimed to investigate the effect of different hepatic inflow interruption methods on bile duct injury. Methods Forty rats were randomly grouped as sham, Pringle maneuver (PM) and hepatic arterial blood flow open (HAFO) groups. We evaluated liver histology and function in liver sections, and biliary histology, cholangiocyte apoptosis and proliferation, cytokine production, and bile composition. RNA sequencing is performed to explore possible molecular mechanisms. The Blood-biliary barrier permeability and tight junctions were analyzed by HRP injection, immunofluorescence staining and analysis of ZO-1 expression by immunoblotting. Results HAFO significantly attenuated ischemia-induced liver injury and decreased ALT, ALP, TBIL, and DBIL levels in serum. The histopathological observations showed that bile duct injury in the PM group was more serious than that in the HAFO group. The numbers of apoptotic biliary epithelial cells in HAFO-treated rat bile ducta were lower than those in the PM group. RNA-seq showed that tight junctions may be related to the mechanism underlying the protective effect of HAFO, as shown by the reduced HRP levels and increased ZO-1 and claudin-1/3 expression in the HAFO group compared to the PM group. Conclusion Compared with PM, HAFO alleviated the ischemic injury to the biliary system, which was characterized by decreased biliary epithelial cell apoptosis, reduced inflammatory responses and decreased blood-biliary-barrier permeability.

Ocular surface and in vivo confocal microscopic findings in patients with active tyhroid eye disease treated with glucocorticoids.

To evaluate the effect of treatment with high-dose glucocorticoids (HDG) on ocular surface parameters and in vivo confocal microscopy (IVCM) findings in patients with active thyroid eye disease (TED). This study included 15 active TED patients treated with a cumulative HDG dose of 4.5g (500 mg and then 250 mg for 6 weeks each). Disease activity was assessed by the clinical activity score (CAS), Graves' Orbitopathy-specific Quality of Life (GO-QoL) questionnaire, ocular surface disease index (OSDI), tear break-up time(T-BUT), ocular surface staining and IVCM. The mean follow-up time was 48.6 months. The treatment resulted in a significant improvement in the T-BUT, ocular surface staining values. In comparison to the baseline, a notable reduction in CAS, OSDI and an increase in GO-QoL were seen after one year. IVCM showed a significant decrease in the number of dendritic cells, activated keratocytes, the degree of tortuosity, there was also a significant increase in the number of basal epithelial cells and total nerves over one-year period. All of those changes remained stable during the follow-up period. HDG treatment results in a sustained decrease in corneal inflammatory cells, an increase in tear film stability, and an improvement in disease symptoms and QoL scores for up to 4 years.

Phenotype of Urine Sediment Cells in Patients with Bladder Cancer

Phenotype of urine sediment cells were studied in patients with bladder cancer depending on the cancer stage and recurrence prognosis. In T1N0M0 stage, the number of lymphocytes decreased, in T2N0M0 stage, the most pronounced shift was an increase in the number of erythrocytes. Irrespectively of the disease stage, we observed increased number of innate immunity cells and cells that inhibit antitumor immunity in the composition of the leukocyte fraction of urine sediment cells. At T1N0M0 stage, the epithelial-endothelial fraction was characterized by increased content of cells expressing CD13 marker (responsible for tumor growth and metastasis) and reduced number of cells expressing CD15 marker (responsible for intercellular adhesion). In patients developing relapse of bladder cancer, the number of lymphocytes was decreased in urine sediment cells and the number of epithelial and endothelial cells expressing CD13 marker was increased.

Dendrobium fimbriatum polysaccharide ameliorates DSS-induced intestinal mucosal injury by IL-22-regulated intestinal stem cell regeneration

Ulcerative colitis (UC) is a chronic inflammatory bowel disease with unknown etiology and difficult treatment. In this study, the intervention effect of Dendrobium fimbriatum Hook polysaccharide (cDFPW1) on UC was verified by constructing a dextran sulfate sodium (DSS)-induced colitis mouse model, and the protective effect of cDFPW1 on intestinal mucosal integrity in UC was explored by the co-culture system consisting of intestinal organoids and lamina propria lymphocytes (LPLs) combined with the experiment of microbial depletion mice. Results showed that cDFPW1 significantly alleviated UC symptoms in mice and promoted the proliferation of intestinal epithelial cells. Importantly, cDFPW1 could directly improve DSS-induced morphological damage of intestinal organoids and increase the number of epithelial cells, which was validated in mice. During repair, an increase in the number of Lgr5+ cells in intestinal organoids and mouse intestines was promoted by cDFPW1. Meanwhile, cDFPW1 promoted intestinal stem cells (ISCs)-mediated intestinal epithelial regeneration by significantly upregulating IL-22 expression. We further confirmed that the secretion of IL-22 was mediated by LPLs. Together, these findings suggest that cDFPW1 promotes ISCs regeneration by LPLs-mediated up-regulation of IL-22 to protect the intestinal mucosal integrity, thereby playing an important role in improving UC.

Interleukin 11 confers resistance to dextran sulfate sodium-induced colitis in mice

Intestinal homeostasis is tightly regulated by epithelial cells, leukocytes, and stromal cells, and its dysregulation is associated with inflammatory bowel diseases. Interleukin (IL)-11, a member of the IL-6 family of cytokines, is produced by inflammatory fibroblasts during acute colitis. However, the role of IL-11 in the development of colitis is still unclear. Herein, we showed that IL-11 ameliorated DSS-induced acute colitis in mouse models. We found that deletion of Il11ra1 or Il11 rendered mice highly susceptible to DSS-induced colitis compared to the respective control mice. The number of apoptotic epithelial cells was increased in DSS-treated Il11ra1- or Il11-deficient mice. Moreover, we showed that IL-11 production was regulated by reactive oxygen species (ROS) produced by lysozyme M-positive myeloid cells. These findings indicate that fibroblast-produced IL-11 plays an important role in protecting the mucosal epithelium in acute colitis. Myeloid cell-derived ROS contribute to the attenuation of colitis through the production of IL-11.

Mechanism underlying the modulation of milk production by incomplete milking.

Mammary gland secretory activity is modulated by systemic and local factors; however, the relationship between these factors is unknown. The aim of this study was to determine how a local factor, such as incomplete milking, affects mammary epithelial cell activity, number, and responsiveness to blood prolactin (PRL). Eight cows in mid-lactation were differentially milked (i.e., their right quarters were milked incompletely at approximately 70%, and their left quarters were milked completely, twice daily for 4 wk). Throughout the experiment, milk yield was measured at the quarter level. Milk samples were collected from each quarter once a week to assess the milk components, and epithelial cell concentrations, as well as to isolate milk fat globule RNA. In the weeks before and after the experiment, mammary gland functional capacity was evaluated by measuring the volume of milk harvested after complete filling of the gland. At the end of the last experimental week, mammary gland biopsies were performed on each rear quarter. The milk production of quarters milked completely remained stable during the treatment period, whereas, as expected, the milk production of quarters milked incompletely was only 53% of completely milked quarters at the end of the period. Accordingly, the expression of genes related to milk synthesis (CSN2, LALBA, and ACACA) in milk fat was lower in the quarters that were milked incompletely. Incomplete milking decreased the milk lactose content, indicating a loss of integrity of tight junctions. The total yield of epithelial cells in milk was not affected, but their concentration in milk, the BAX:BCL2 gene expression ratio, and the loss of mammary functional capacity were greater in the quarters milked incompletely, suggesting an acceleration of involution in those quarters. The expression of the short isoform of the PRL receptor gene (PRLR) tended to be lower, and the expression of STAT5A and STAT5B tended to decline in the quarters milked incompletely. In mammary gland biopsy samples, the number of both short and long isoforms of the PRLR were not affected, nor were the amount and activation of STAT3 and STAT5. However, the ratio of PRLR short isoform to PRLR long isoform was lower in the quarters milked incompletely. The decrease in milk yield induced by incomplete milking is rapid and associated with a decrease in mammary epithelial cell activity and a decrease in the number of secretory epithelial cells. The results of this experiment provide only limited support for the hypothesis that modulation of the mammary gland's responsiveness to PRL is part of the mechanism by which local factors, such as incomplete milking, modulate milk synthesis.

Epididymis cell atlas in a patient with a sex development disorder and a novel NR5A1 gene mutation.

This study aims to characterize the cell atlas of the epididymis derived from a 46,XY disorders of sex development (DSD) patient with a novel heterozygous mutation of the nuclear receptor subfamily 5 group A member 1 (NR5A1) gene. Next-generation sequencing found a heterozygous c.124C>G mutation in NR5A1 that resulted in a p.Q42E missense mutation in the conserved DNA-binding domain of NR5A1. The patient demonstrated feminization of external genitalia and Tanner stage 1 breast development. The surgical procedure revealed a morphologically normal epididymis and vas deferens but a dysplastic testis. Microfluidic-based single-cell RNA sequencing (scRNA-seq) analysis found that the fibroblast cells were significantly increased (approximately 46.5%), whereas the number of main epididymal epithelial cells (approximately 9.2%), such as principal cells and basal cells, was dramatically decreased. Bioinformatics analysis of cell-cell communications and gene regulatory networks at the single-cell level inferred that epididymal epithelial cell loss and fibroblast occupation are associated with the epithelial-to-mesenchymal transition (EMT) process. The present study provides a cell atlas of the epididymis of a patient with 46,XY DSD and serves as an important resource for understanding the pathophysiology of DSD.

Analogy between the oral microbial flora found at the level of carious lesions, endodontic spaces and prosthetic components

Nowadays, eating habits and lifestyles are moving towards a diet high in sugars and carbohydrates, which is extremely harmful to both oral integrity and systemic health. The aim of the study was to identify the variations between the bacterial plaque formed in carious lesions, infected root canals and prosthetic components, in comparison with commensal oral microflora. The study was carried out on a group of 21 patients aged between 7 and 84 years, 13 women and 8 men, with various oral diseases, which were diagnosed during the specialized examination. The preparation of the medical record, the signing of the informed consent, the anamnesis and the consultation were carried out in the private dental office where the patients presented for a personalized treatment plan, and the examination of the pathological products and the interpretation of the results obtained in the Microbiology Discipline, “Carol Davila” University of Medicine and Pharmacy, Bucharest, Romania. The study carried out on the preparations taken from patients with carious lesions reveals the presence of numerous G+ cocci arranged in short chains (23%), frequent G+ cocci in diplo, thin G+ bacilli with rounded ends and G+ cocci in diplo, arranged in small piles (15 %), and values lower than 8% are represented by G+ in diplo cocci, lanceolate, rare G+ in diplo cocci, very rare fusiform bacilli and dentinal fragments. The study carried out on the preparations taken from the endodontic space showed a remarkable percentage of epithelial cells and leukocytes (28%), rare fusiform bacilli, very small fragments of epithelial tissue, thin G+ bacilli with rounded ends, G+ cocci in diplo and in small piles and buns arranged in short chains (5-6%). The study carried out on the preparations taken from the level of prosthetic components, recorded a relatively high number of epithelial cells (19%), G+ cocci arranged in diplo, lanceolate and in small piles (13%), thin G+ bacilli, with rounded ends (10 %), cocci in short chains (10%) and rare polymorphonuclear leukocytes (10%), while at the opposite pole, squamous epithelial cells, fusobacteria and pleomorphic forms (3%) are found.

Oral immunization of mice with recombinant Lactobacillus plantarum expressing a Trichinella spiralis galectin induces an immune protection against larval challenge

BackgroundTrichinella spiralis is an important foodborne parasite that presents a severe threat to food safety. The development of an anti-Trichinella vaccine is an important step towards controlling Trichinella infection in food animals and thus ensure meat safety. Trichinella spiralis galectin (Tsgal) is a novel protein that has been identified on the surface of this nematode. Recombinant Tsgal (rTsgal) was found to participate in larval invasion of intestinal epithelium cells (IECs), whereas anti-rTsgal antibodies impeded the invasion.MethodsThe rTsgal/pSIP409- pgsA′ plasmid was constructed and transferred into Lactobacillus plantarum strain NC8, following which the in vitro biological properties of rTsgal/NC8 were determined. Five groups of mice were orally immunized three times, with a 2-week interval between immunizations, with recombinant NC8-Tsgal, recombinant NC8-Tsgal + α-lactose, empty NC8, α-lactose only or phosphate-buffered saline (PBS), respectively. The vaccinated mice were infected orally with T. spiralis larvae 2 weeks following the last vaccination. Systemic and intestinal local mucosal immune responses and protection were also assessed, as were pathological changes in murine intestine and skeletal muscle.ResultsrTsgal was expressed on the surface of NC8-Tsgal. Oral immunization of mice with rTsgal vaccine induced specific forms of serum immunoglobulin G (IgG), namely IgG1/IgG2a, as well as IgA and gut mucosal secretion IgA (sIgA). The levels of interferon gamma and interleukin-4 secreted by cells of the spleen, mesenteric lymph nodes, Peyer's patches and intestinal lamina propria were significantly elevated at 2–6 weeks after immunization, and continued to rise following challenge. Immunization of mice with the oral rTsgal vaccine produced a significant immune protection against T. spiralis challenge, as demonstrated by a 57.28% reduction in the intestinal adult worm burden and a 53.30% reduction in muscle larval burden, compared to the PBS control group. Immunization with oral rTsgal vaccine also ameliorated intestinal inflammation, as demonstrated by a distinct reduction in the number of gut epithelial goblet cells and mucin 2 expression level in T. spiralis-infected mice. Oral administration of lactose alone also reduced adult worm and larval burdens and relieved partially inflammation of intestine and muscles.ConclusionsImmunization with oral rTsgal vaccine triggered an obvious gut local mucosal sIgA response and specific systemic Th1/Th2 immune response, as well as an evident protective immunity against T. spiralis challenge. Oral rTsgal vaccine provided a prospective approach for control of T. spiralis infection.Graphical

The spring-like effect of microRNA-31 in balancing inflammatory and regenerative responses in colitis.

Inflammatory bowel diseases (IBDs) are chronic inflammatory disorders caused by the disruption of immune tolerance to the gut microbiota. MicroRNA-31 (MIR31) has been proven to be up-regulated in intestinal tissues from patients with IBDs and colitis-associated neoplasias. While the functional role of MIR31 in colitis and related diseases remain elusive. Combining mathematical modeling and experimental analysis, we systematically explored the regulatory mechanism of MIR31 in inflammatory and epithelial regeneration responses in colitis. Level of MIR31 presents an "adaptation" behavior in dextran sulfate sodium (DSS)-induced colitis, and the similar behavior is also observed for the key cytokines of p65 and STAT3. Simulation analysis predicts MIR31 suppresses the activation of p65 and STAT3 but accelerates the recovery of epithelia in colitis, which are validated by our experimental observations. Further analysis reveals that the number of proliferative epithelial cells, which characterizes the inflammatory process and the recovery of epithelia in colitis, is mainly determined by the inhibition of MIR31 on IL17RA. MIR31 promotes epithelial regeneration in low levels of DSS-induced colitis but inhibits inflammation with high DSS levels, which is dominated by the competition for MIR31 to either inhibit inflammation or promote epithelial regeneration by binding to different targets. The binding probability determines the functional transformation of MIR31, but the functional strength is determined by MIR31 levels. Thus, the role of MIR31 in the inflammatory response can be described as the "spring-like effect," where DSS, MIR31 action strength, and proliferative epithelial cell number are regarded as external force, intrinsic spring force, and spring length, respectively. Overall, our study uncovers the vital roles of MIR31 in balancing inflammation and the recovery of epithelia in colitis, providing potential clues for the development of therapeutic targets in drug design.

From morphological to ecological adaptation of the cornea in Oxudercinae fishes

The outer cornea plays an important role in animal adaptation and survival in different environments. however, research on the morphological and ecological adaptation of corneal structure in amphibious fishes is limited. In this study, scanning electron microscopy (SEM) was used to evaluate the microstructure and adaptation of corneal epithelial cells in Oxudercinae. The results showed that the corneas of Oxudercinae species possess microridges, microvilli, and microplicae, as well as different numbers of epithelial cells. The morphological structure of corneal epithelial cells, observed by collecting samples and comparing the results with previous results, also showed different adaptive characteristics for moving between water and land. Further analyses revealed significant differences in epithelial cell density (F4, 22 = 5.436, P=0.003) and microridge width (F4, 22 = 8.392, P&amp;lt;0.001) among species with different levels of aquatic dependence. In addition, significant negative correlations of epithelial cell density with microridge width and separation width were confirmed (P&amp;lt;0.05). Interestingly, significant negative correlations of habitat type with cell density and microridges were uncovered, as well as a positive correlation between habitat type and separation width (P&amp;lt;0.05). The results indicated that the corneal structure of Oxudercinae species has characteristics of adaptation to an amphibious lifestyle.