Epithelial Specimens Research Articles

Isolation, molecular characterization, and genetic diversity of recently isolated foot-and-mouth disease virus serotype A in Egypt.

Foot-and-mouth Disease (FMD) is a highly contagious viral disease affecting all hoof-cloven animals. Serotypes A, O and SAT 2 of the foot-and-mouth disease virus (FMDV) are circulating in Egypt. The present study aimed to identify and molecularly characterize the FMDV strains circulating in Northern Egypt during an epidemic that struck the nation in 2022. RNA was extracted from the epithelial specimens, vesicular fluid from affected cattle. The samples were screened using real-time reverse-transcription polymerase chain reaction (RT-PCR) targeting the RNA-dependent RNA polymerase (RdRp) gene. Positive samples underwent individual serotype-specific amplification using primers designed for VP1 of O, A, and SAT 2 serotypes. Subsequently, direct sequencing was performed on the positive samples. The real-time RT-PCR detected positive samples from epithelial and vesicular fluid samples, but not in the blood of infected animals. Out of the 16 samples, seven tested positive for FMDV serotype A. Of these seven positive samples, six were categorized as serotype A-African topotype-G-IV, and these positive samples were isolated in BHK-21 cells, yielding an overt cytopathic effect caused by the virus. In conclusion, it is necessary to sustain continuous surveillance of the evolution of circulating FMDV strains to facilitate the assessment and aid in the selection of vaccine strains for the effective control of FMDV in Egypt.

High hsa_circ_0081621 expression indicates a poor prognosis of laryngeal squamous cell carcinoma: A bioinformatics analysis and retrospective clinical study.

Laryngeal squamous cell carcinoma (LSCC) is the second most common malignant tumour of the head and neck with a low 5-year survival rate. There is need to identify novel biomarkers for diagnosis and treatment of LSCC. The present study identified differentially expressed circular RNAs (circRNAs/circs) in LSCC and larynx adjacent non-carcinoma epithelial specimens by analysing the circRNA microarray dataset GSE117001. hsa_circ_0081621 had highest expression among three circRNAs (hsa_circ_0015211, hsa_circ_0023326 and hsa_circ_0081621) in LSCC specimens by reverse transcription-quantitative PCR. The expression levels of hsa_circ_0081621 in 67 LSCC specimens were detected by fluorescence in situ hybridization (FISH). Expression levels of hsa_circ_0081621 were analysed in relation to clinicopathological parameters and prognosis of patients with LSCC. According to FISH results, 59.7% of LSCC specimens exhibited high hsa_circ_0081621 expression. In LSCC specimens, hsa_circ_0081621 high expression was associated with lymph node metastasis and high clinical stage. High expression levels of hsa_circ_0081621 were associated with a poor 5-year overall survival rate in patients with LSCC. In addition, high hsa_circ_0081621 expression was an independent prognostic factor for patients with LSCC. hsa_circ_0081621 may participate in malignant progression of LSCC, and its high expression could be used for prognostic assessment of patients with LSCC.

HPV prevalence and genotype distribution among 38 056 women in Weifang, China: a cross-sectional study

ObjectivesHuman papillomavirus (HPV) infection is closely associated with cervical cancer, especially the persistent infection of high-risk HPV (HR-HPV) genotypes. Therefore, investigating the HPV prevalence, age-specific, genotype distribution and the impact...

PCB: A pseudotemporal causality-based Bayesian approach to identify EMT-associated regulatory relationships of AS events and RBPs during breast cancer progression

During breast cancer metastasis, the developmental process epithelial-mesenchymal (EM) transition is abnormally activated. Transcriptional regulatory networks controlling EM transition are well-studied; however, alternative RNA splicing also plays a critical regulatory role during this process. Alternative splicing was proved to control the EM transition process, and RNA-binding proteins were determined to regulate alternative splicing. A comprehensive understanding of alternative splicing and the RNA-binding proteins that regulate it during EM transition and their dynamic impact on breast cancer remains largely unknown. To accurately study the dynamic regulatory relationships, time-series data of the EM transition process are essential. However, only cross-sectional data of epithelial and mesenchymal specimens are available. Therefore, we developed a pseudotemporal causality-based Bayesian (PCB) approach to infer the dynamic regulatory relationships between alternative splicing events and RNA-binding proteins. Our study sheds light on facilitating the regulatory network-based approach to identify key RNA-binding proteins or target alternative splicing events for the diagnosis or treatment of cancers. The data and code for PCB are available at: http://hkumath.hku.hk/~wkc/PCB(data+code).zip.

Pericytes may facilitate SARS-CoV-2 entry into the nervous system

Objective: Although the neurological and olfactory symptoms of coronavirus disease 2019 have been identified, the neurotropic properties of the causative virus, severe acute respiratory syndrome-associated coronavirus 2 (SARS-CoV-2), remain unknown. We sought to identify the susceptible cell types and potential routes of SARS-CoV-2 entry into the central nervous system, olfactory system, and respiratory system. Methods: We collected single-cell RNA data from normal brain and nasal epithelium specimens, along with bronchial, tracheal, and lung specimens in public datasets. The susceptible cell types that express SARS-CoV-2 entry genes were identified using single-cell RNA sequencing and the expression of the key genes at protein levels was verified by immunohistochemistry. We compared the coexpression patterns of the entry receptor angiotensin-converting enzyme 2 (ACE2) and the spike protein priming enzyme transmembrane serine protease (TMPRSS)/cathepsin L among the specimens. Results: The SARS-CoV-2 entry receptor ACE2 and the spike protein priming enzyme TMPRSS/cathepsin L were coexpressed by pericytes in brain tissue; this coexpression was confirmed by immunohistochemistry. In the nasal epithelium, ciliated cells and sustentacular cells exhibited strong coexpression of ACE2 and TMPRSS. Neurons and glia in the brain and nasal epithelium did not exhibit coexpression of ACE2 and TMPRSS. However, coexpression was present in ciliated cells, vascular smooth muscle cells, and fibroblasts in tracheal tissue; ciliated cells and goblet cells in bronchial tissue; and alveolar epithelium type 1 cells, AT2 cells, and ciliated cells in lung tissue. Conclusion: Neurological symptoms in patients with coronavirus disease 2019 could be associated with SARS-CoV-2 invasion across the blood–brain barrier via pericytes. Additionally, SARS-CoV-2–induced olfactory disorders could be the result of localized cell damage in the nasal epithelium.

Resveratrol inhibits development of colorectal adenoma via suppression of LEF1; comprehensive analysis with connectivity map.

Although many chemopreventive studies on colorectal tumors have been reported, no effective and safe preventive agent is currently available. We searched for candidate preventive compounds against colorectal tumor comprehensively from United States Food and Drug Administration (FDA)-approved compounds by using connectivity map (CMAP) analysis coupled with in vitro screening with colorectal adenoma (CRA) patient-derived organoids (PDOs). We generated CRA-specific gene signatures based on the DNA microarray analysis of CRA and normal epithelial specimens, applied them to CMAP analysis with 1309 FDA-approved compounds, and identified 121 candidate compounds that should cancel the gene signatures. We narrowed them down to 15 compounds, and evaluated their inhibitory effects on the growth of CRA-PDOs in vitro. We finally identified resveratrol, one of the polyphenolic phytochemicals, as a compound showing the strongest inhibitory effect on the growth of CRA-PDOs compared with normal epithelial PDOs. When resveratrol was administered to ApcMin/+ mice at 15 or 30 mg/kg, the number of polyps (adenomas) was significantly reduced in both groups compared with control mice. Similarly, the number of polyps (adenomas) was significantly reduced in azoxymethane-injected rats treated with 10 or 100 mg/resveratrol compared with control rats. Microarray analysis of adenomas from resveratrol-treated rats revealed the highest change (downregulation) in expression of LEF1, a key molecule in the Wnt signaling pathway. Treatment with resveratrol significantly downregulated the Wnt-target gene (MYC) in CRA-PDOs. Our data demonstrated that resveratrol can be the most effective compound for chemoprevention of colorectal tumors, the efficacy of which is mediated through suppression of LEF1 expression in the Wnt signaling pathway.

Identifying BRCA wild-type ovarian cancers with a homologous recombination deficiency phenotype (101)

<b>Objectives:</b> It is unknown how best to evaluate <i>BRCA</i> wild-type ovarian carcinomas for homologous recombination deficiency (HRD), and previous studies did not show a survival advantage for <i>BRCA1</i> methylation. We sought to further characterize <i>BRCA1/RAD51C</i> promoter methylation in ovarian cancer (OC). We hypothesized that a combined assay with quantitative methylation and mutation assessment would identify <i>BRCA</i> wild-type ovarian carcinomas with an HRD phenotype. <b>Methods:</b> Digital droplet PCR primers and probes were designed to target CpG sites in the promoter regions of <i>BRCA1</i> and <i>RAD51C</i> known to be associated with transcriptional repression. Multiple aspects of the assay were tested and optimized in a CLIA environment. The fraction of methylated alleles was corrected for neoplastic cellularity and ≥60% was considered highly methylated. The methylated ddPCR (MddPCR) assay was then applied to DNA from banked epithelial ovarian cancer specimens previously assessed by methylation-sensitive PCR (MSP) and BROCA next-generation sequencing. RNA expression of a subset of <i>BRCA1</i> methylated samples was determined using RNA-seq, and expression was compared with methylation using LOESS regression. Clinical characteristics were then compared between highly methylated and unmethylated samples using a t-test for continuous variables, Fisher's exact test for categorical variables, and the log-rank test for overall survival (OS). Additional cases with germline/somatic mutations in homologous recombination repair (HRR) genes identified by BROCA sequencing were included in an overall survival analysis using the log-rank test. <b>Results:</b> Total 370 epithelial OC specimens were tested by MddPCR. MddPCR had 100% concordance with MSP. Among the samples, 9.5% were positive for <i>BRCA1</i> promoter methylation, and 3% were positive for <i>RAD51C</i> promoter methylation ≥5%. RNA expression analysis of a sub-set of <i>BRCA1</i> methylated samples demonstrated that the degree of <i>BRCA1</i> methylation was negatively correlated with <i>BRCA1</i> expression levels (r=-0.82, p<0.001). There were 26 highly methylated neoplastic <i>BRCA1</i> or <i>RAD51C</i> samples, 14 samples with low methylation (5-59% methylated alleles), and 294 samples with no methylation or HRR mutation. The median overall survival was 76 months for highly methylated cases compared to 50 months for unmethylated cases (HR: 0.66, p=0.20). Highly methylated samples were significantly less likely to have had prior chemotherapy exposure (12% vs 31%, <i>p</i>=0.04). Paired primary/recurrent samples were evaluated, two of which contained a highly methylated initial sample and showed a decrease in methylation at the time of recurrence. In the expanded cohort including HRR mutated OC, median OS was 80 months in cases with either high methylation or a non-<i>BRCA</i> HRR mutation compared to 50 months in unmethylated wild-type samples (HR: 0.63, p=0.02), with a survival curve similar to <i>BRCA</i> mutated samples (median OS: 66 months, HR: 0.76, p=0.01). <b>Conclusions:</b> Ovarian cancer that was either highly methylated or had a non-<i>BRCA</i> HRR mutation was associated with increased OS compared to unmethylated/wild-type cases, suggesting an HRD phenotype. Methylated samples had similar OS to HRR mutated cancers; however, survival analysis was limited by the low total number of highly methylated samples. Exposure to chemotherapy may decrease promoter methylation, and analysis of additional paired samples is ongoing. Quantitative methylation analysis could augment tumor sequencing in identifying OC with a favorable response to PARP inhibitors.Fig. 1

Barrier Perturbation in Porcine Peyer's Patches by Tumor Necrosis Factor is Associated With a Dysregulation of Claudins.

The proinflammatory cytokine tumor necrosis factor (TNF) has been described as one of the main mediators of intestinal inflammatory diseases, affecting the composition of tight junction (TJ) proteins and leading to a disruption of the epithelial barrier. An intact intestinal barrier is mandatory, because the follicle-associated epithelium of Peyer’s patches represents the first defense line of the intestinal immune system and ensures a controlled uptake of antigens from the gut lumen. In the current study, we have analyzed the detailed effects of TNF on the follicle-associated epithelium of porcine Peyer’s patches by applying the Ussing chamber technique. Epithelial tissue specimens of Peyer’s patches and the surrounding villus epithelium were mounted into conventional Ussing chambers and incubated with TNF for 10 h. The transepithelial resistance, representing epithelial barrier function of the tissue, was recorded. A reduction of transepithelial resistance was detected after 8 h in Peyer’s patch tissue specimens, whereas the villus epithelium was not significantly affected by TNF. Subsequent molecular analysis of TJ protein expression revealed a marked decrease of claudin-1 and -4, and an increase of claudin-2. In neighboring villus epithelium, no significant changes in the expression of TJ proteins could be shown. A strong increase of TNF receptor-2 (TNFR-2) could also be detected in Peyer’s patches, in agreement with the major role of this receptor in Peyer’s patches. Our findings were in accordance with changes detected by confocal laser scanning immunofluorescence microscopy. The regulation of TNF effects via myosin light chain kinase (MLCK) was analyzed in blocking experiments. Our detailed analysis is the first to show that TNF affects the barrier function of the follicle-associated epithelium of porcine Peyer’s patches but has no effects on the villus epithelium. These findings reveal not only the basic differences of epithelial barrier function between the two structures, but also the significance of Peyer’s patches as a primary mucosal immune defense.

Analysis of Expression of Myofibroblasts in Oral Submucous Fibrosis: An Immunohistochemistry Study

Background: Oral submucous fibrosis (OSMF) is a chronic disease that produces tissue fibrosis and is considered to be a potentially malignant disorder. The exact pathogenesis and malignant conversion mechanism of this disorder are still unknown. Myofibroblasts have been implicated as one of the possible pathological mechanisms responsible for the pathophysiology of OSMF. The present study was conducted to evaluate the expression of myofibroblasts (MF) in normal mucosa and different grades of OSMF. Materials &amp; Methods: The sample consisted of a total of 80 specimens. The study group included specimens from clinically and histopathologically confirmed OSMF patients. The specimens were divided into four groups. Group 1 consisted of 19 specimens of grade III OSMF. Group II had 20 specimens of grade II OSMF, Group 3 with 21 specimens of grade I OSMF, and Group 4 constituted a control group of 20 normal epithelium specimens. Two sections each from all the four groups were obtained. While one section was stained with H and E, the other section was stained immunohistochemically using α-smooth muscle antibody. For analysis, the expression of myofibroblasts was categorized as strong, moderate, weak, or absent. All the results were recorded and subjected to statistical analysis. Results: In OSMF patients, irrespective of the grade, the expression of myofibroblast was strong in 28.33 percent of the patients, while it was moderate and weak in 30.00 percent and 40.00 percent of the patients, respectively. Expression of myofibroblast was noted to be significantly increased in grade III OSMF patients as compared to controls as well as grade I OSMF patients (p-value &lt;0.05). Conclusion: Myofibroblasts expression is significantly raised in OSMF patients. The expression can also be correlated within different grades of OSMF where advanced stages show comparatively high expression of these smooth muscles like fibroblasts. Hence, we suggest that myofibroblasts could be assessed as markers for analyzing the progression of OSMF.

Expression analysis and function of mitochondrial genome-encoded microRNAs

MicroRNAs (miRNAs) play a significant role in nuclear and mitochondrial anterograde and retrograde signaling. Most of the miRNAs found inside mitochondria are encoded in the nuclear genome, with a few mitochondrial genome-encoded non-coding RNAs having been reported. In this study, we have identified 13 mitochondrial genome-encoded microRNAs (mitomiRs), which were differentially expressed in breast cancer cell lines (MCF-7, MDA-MB-468 and MDA-MB-231), non-malignant breast epithelial cell line (MCF-10A), and normal and breast cancer tissue specimens. We found that mitochondrial DNA (mtDNA) depletion and inhibition of mitochondrial transcription led to reduced expression of mitomiRs in breast cancer cells. MitomiRs physically interacted with Ago2, an RNA-induced silencing complex (RISC) protein, in the cytoplasm and inside mitochondria. MitomiRs regulate the expression of both nuclear and mitochondrial transcripts in breast cancer cells. We showed that mitomiR-5 targets the PPARGC1A gene and regulates mtDNA copy number in breast cancer cells. MitomiRs identified in the present study may be a promising tool for expression and functional analysis in patients with a defective mitochondrial phenotype, including cancer and metabolic syndromes. This article has an associated First Person interview with the first author of the paper.

Expression Analysis of Ermin and Listerin E3 Ubiquitin Protein Ligase 1 Genes in the Periphery of Patients with Schizophrenia.

Schizophrenia (SCZ) is a severe mental disorder with an unknown etiology. Recent researches indicate that correct myelination and translational regulation play a role in the pathogeny of SCZ. This study evaluated the expression pattern of Ermin (ERMN) and Listerin E3 ubiquitin protein ligase 1 (LTN1) genes, which play a role in myelination and ribosome quality control, respectively. The expression of the ERMN and LTN1 genes in the peripheral blood (PB) of 50 SCZ patients (male/female: 22/28, age (mean ± standard deviation (SD)): 35.9 ± 5.6) and 50 matched healthy controls (male/female: 23/27, age (mean ± SD): 34.7 ± 5.4) were assessed using quantitative polymerase chain reaction. Additionally, we used a bioinformatics approach based on microarray dataset analysis to examine the expression of these two genes in olfactory epithelium (OE) specimens. The expression of ERMN demonstrated no significant differences in PB samples among SCZ patients and healthy controls (adjusted P-value = 0.101). The expression of LTN1 was significantly higher in PB samples obtained from female patients compared with sex-matched controls (posterior beta = 1.734, adjusted P-value < 0.0001). Significant correlations were found between expression of the mentioned genes in PB samples both among SCZ patients and among healthy controls (r = 0.485, P < 0.001 and r = 0.516, P < 0.001, respectively). According to our in silico findings, the ERMN expression levels in OE samples of SCZ were statistically higher than those in controls (log2FC = 1.93, adj.P.Val = 9.66E-15). On the contrary, LTN1 expression levels in OE samples were statistically lower in SCZ cases versus controls (log2FC = - 0.77, adj.P.Val = 2.14E-06). Besides, a significant correlation was found between the expression of the mentioned genes in OE samples (r = - 0.60, P < 0.001). In conclusion, the present study is the first evidence to highlight the expression of the ERMN and LTN1 genes in the periphery of SCZ patients. Our findings may provide light on the SCZ's pathogeny.

Immunohistochemical Expression of Epithelial Mesenchymal Transition Proteins in Squamous Cell Carcinoma of the Oral Cavity

Background & Objective: Epithelial-Mesenchymal transition (EMT) is known to be a possible mechanism in tumor progression; however, there is insufficient evidence to support the contribution of this process in human cancers. The present study aimed to evaluate the expression of EMT markers in normal oral epithelium and oral squamous cell carcinoma and also correlates with some clinicopathological parameters.Methods: This study was conducted on 70 samples, including 20 cases of normal epithelium and 50 cases of Oral Squamous cell Carcinoma (OSCC). To examine the expression level of these proteins, immunohistochemical staining was performed for samples using E-cadherin and N-cadherin monoclonal antibodies.Results: Reduced expression of E-cadherin was observed in 74% of OSCC and 15% of normal epithelium samples; this difference was statistically significant (P˂0.000). With the progression of SCC from well towards poor differentiation, the E-cadherin expression decreased; however, this difference was not statistically significant (P=0.642). Normal epithelial specimens were negative for N-cadherin expression in 75% of cases, whereas OSCC specimens showed high expression of N-cadherin in 46% of cases, this difference was statistically significant (P=0.01). Although 62.5% of poorly differentiated OSCC showed high expression of N-cadherin, the difference between the histopathological grades was not significant (P=0.586). No significant relationship was found between markers expression and patient’s age, gender, and tumor location.Conclusion: This study showed that OSCC tissues showed high EMT phenotype (reduced E-cadherin expression and high expression of N-cadherin) compared to normal oral mucosa which may indicate the possible key role of EMT mechanism during oral carcinogenesis.

Construction autophagy-related prognostic risk signature to facilitate survival prediction, individual treatment and biomarker excavation of epithelial ovarian cancer patients

BackgroundExisting clinical methods for prognosis evaluating for Epithelial Ovarian Cancer (EOC) patients had defects of invasive, unsystematic and subjective and little data are available for individualizing treatment, therefore, to identify potential prognostic markers and new therapeutic targets for EOC is urgently required.ResultsExpression of 232 autophagy-related genes (ARGs) in 354 EOC and 56 human ovarian surface epithelial specimens from 7 independent laboratories were analyzed, 31 mRNAs were identified as DEARGs. We did functional and pathway enrichment analysis and constructed protein–protein interaction network for all DEARGs. To screen out candidate DEARGs related to EOC patients’ survival and construct an autophagy-related prognostic risk signature, univariate and multivariate Cox proportional hazards models were established separately. Finally, 5 optimal independent prognostic DEARGs (PEX3, DNAJB9, RB1, HSP90AB1 and CXCR4) were confirmed and the autophagy-related risk model was established by the 5 prognostic DEARGs. The accuracy and robustness of the prognostic risk model for survival prediction were evaluated and verified by analyzing the correlation between EOC patients’ survival status, clinicopathological features and risk scores.ConclusionsThe autophagy-related prognostic risk model can be independently used to predict overall survival in EOC patients, it can also potentially assist in individualizing treatment and biomarker development.

Par3/integrin β1 regulates embryo adhesion via changing endometrial luminal epithelium polarity†.

The objective is to investigate the pathophysiological significance of Par3 and integrin β1 with regard to the functionality of the endometrial luminal epithelium (LE). Design: laboratory study; setting: university research laboratory. Analysis involved endometrial aspirates and endometrial adenocarcinoma cells (HEC-1A) and endometrial carcinoma cells (RL95-2). We first examined the expression and localization of Par3 and integrin β1 in HEC-1A cells and RL95-2 cells. Then we knocked down Par3 and integrin β1 in HEC-1A cells and RL95-2 cells, respectively, and found that Par3/integrin β1 affected embryo adhesion by regulating the intercellular tight junctions' (TJs') structure and thus the polarity of the endometrial LE. These findings were also confirmed in the endometrium specimens from human and mice. The main outcome measures were the expression and localization of Par3 and integrin β1 in the endometrial epithelial cell lines and endometrium specimens and the regulations of Par3 and integrin β1 on TJs, polarity, and embryo adhesion. Following the knockdown of Par3 in HEC-1A cells, there was a reduction in the complexity of the TJs and cell polarity, and the adhered blastocysts number was significantly increased. However, the reduction of integrin β1 in RL95-2 cells resulted in effects that directly opposed those following the knockdown of Par3 in HEC-1A cells. Estrogen and progesterone reduced the expression of Par3 and promoted the expression of integrin β1 in HEC-1A cells. Par3/integrin β1 regulates embryo adhesion by regulating intercellular TJs' structure and polarity of endometrial LE under the action of ovarian hormones.

Integrin-linked kinase pathway in heterogeneous pulmonary sarcomatoid carcinoma.

Pulmonary sarcomatoid carcinoma (PSC) is classified as poorly differentiated, and non-small cell lung carcinomas that contained a component of sarcoma or sarcoma-like differentiation are rare. The underlying carcinogenetic mechanism governing PSC remains unclear. The current study investigated the underlying carcinogenetic mechanism of PSC based on the hypothesis that it involves the epithelial-mesenchymal transition (EMT) process. Mutation analysis of PSCs, including carcinosarcoma, pleomorphic carcinoma and epithelial carcinoma specimens, was performed using targeted deep sequencing, whole transcriptome analysis and digital spatial profiling (DSP). PSCs exhibit a distinct mutation profile, with TP53, SYNE1 and APC mutations. Therefore, clustering of the gene expression profiles allowed the PSCs to be distinguished from the epithelial carcinomas. Increased gene expression of fibronectin in PSC was an important contributor to differential profiles. Pathway analysis revealed enhanced activity of the integrin-linked kinase (ILK) signaling pathway in the PSCs. DSP analysis using 56 antibodies of marker proteins confirmed significantly higher expression of fibronectin in PSCs. Intratumor heterogeneity of fibronectin expression was observed in sarcoma components. In conclusion, epithelial-mesenchymal transition process mediated by ILK signaling may be associated with carcinogenetic mechanisms of PSC. Overexpression of fibronectin mediated by ILK signaling appears to serve a role in the EMT involved in the PSC transformation process.

A novel evaluation method for Ki-67 immunostaining in paraffin-embedded tissues.

The Ki-67 labeling index is traditionally used to investigate tumor aggressiveness. However, no diagnostic or prognostic value has been associated to the heterogeneous pattern of nuclear positivity. The aims of this study were to develop a classification for the patterns of Ki-67-positive nuclei; to search scientific evidence for the Ki-67 expression and location throughout the cell cycle; and to develop a protocol to apply the classification of patterns of Ki-67-positive nuclei in squamous epithelium with different proliferative activities. Based on empirical observation of paraffin sections submitted to immunohistochemistry for the determination of Ki-67 labeling index and literature review about Ki-67 expression, we created a classification of the patterns of nuclear positivity (NP1, NP2, NP3, NP4, and mitosis). A semi-automatic protocol was developed to identify and quantify the Ki-67 immunostaining patterns in target tissues. Two observers evaluated 7000 nuclei twice to test the intraobserver reliability, and six evaluated 1000 nuclei to the interobserver evaluation. The results showed that the immunohistochemical patterns of Ki-67 are similar in the tumoral and non-tumoral epithelium and were classified without difficulty. There was a high intraobserver reliability (Spearman correlation coefficient > 0.9) and moderate interobserver agreement (k = 0.523). Statistical analysis showed that non-malignant epithelial specimens presented a higher number of NP1 (geographic tongue = 83.8 ± 21.8; no lesion = 107.6 ± 52.7; and mild dysplasia = 86.6 ± 25.8) when compared to carcinoma in Situ (46.8 ± 34.8) and invasive carcinoma (72.6 ± 37.9). The statistical evaluation showed significant difference (p < 0.05). Thus, we propose a new way to evaluate Ki-67, where the pattern of its expression may be associated with the dynamics of the cell cycle. Future proof of this association will validate the use of the classification for its possible impact on cancer prognosis and guidance on personalized therapy.

Immunohistochemical Expression of Epithelial Mesenchymal Transition Proteins in Squamous Cell Carcinoma of the Oral Cavity.

Background & Objective:Epithelial-Mesenchymal transition (EMT) is known to be a possible mechanism in tumor progression; however, there is insufficient evidence to support the contribution of this process in human cancers. The present study aimed to evaluate the expression of EMT markers in normal oral epithelium and oral squamous cell carcinoma and also correlates with some clinicopathological parameters.Methods:This study was conducted on 70 samples, including 20 cases of normal epithelium and 50 cases of Oral Squamous cell Carcinoma (OSCC). To examine the expression level of these proteins, immunohistochemical staining was performed for samples using E-cadherin and N-cadherin monoclonal antibodies.Results:Reduced expression of E-cadherin was observed in 74% of OSCC and 15% of normal epithelium samples; this difference was statistically significant (P˂0.000). With the progression of SCC from well towards poor differentiation, the E-cadherin expression decreased; however, this difference was not statistically significant (P=0.642). Normal epithelial specimens were negative for N-cadherin expression in 75% of cases, whereas OSCC specimens showed high expression of N-cadherin in 46% of cases, this difference was statistically significant (P=0.01). Although 62.5% of poorly differentiated OSCC showed high expression of N-cadherin, the difference between the histopathological grades was not significant (P=0.586). No significant relationship was found between markers expression and patient’s age, gender, and tumor location. Conclusion:This study showed that OSCC tissues showed high EMT phenotype (reduced E-cadherin expression and high expression of N-cadherin) compared to normal oral mucosa which may indicate the possible key role of EMT mechanism during oral carcinogenesis.

Uterine Epithelial Progesterone Receptor Governs Uterine Receptivity Through Epithelial Cell Differentiation.

Progesterone receptor (PGR) is indispensable for pregnancy in mammals. Uterine PGR responds to the heightened levels of ovarian progesterone (P4) after ovulation and regulates uterine gene transcription for successful embryo implantation. Although epithelial and stromal P4-PGR signaling may interact with each other to form appropriate endometrial milieu for uterine receptivity and the subsequent embryo attachment, it remains unclear what the specific roles of epithelial P4-PGR signaling in the adult uterus are. Here we generated mice with epithelial deletion of Pgr in the adult uterus (Pgrfl/flLtfCre/+ mice) by crossing Pgr-floxed and Ltf-Cre mice. Pgrfl/flLtfCre/+ mice are infertile due to the impairment of embryo attachment. Pgrfl/flLtfCre/+ uteri did not exhibit epithelial growth arrest, suggesting compromised uterine receptivity. Both epithelial and stromal expressions of P4-responsive genes decreased in Pgrfl/flLtfCre/+ mice during the peri-implantation period, indicating that epithelial Pgr deletion affects not only epithelial but stromal P4 responsiveness. In addition, uterine LIF, an inducer of embryo attachment, was decreased in Pgrfl/flLtfCre/+ mice. The RNA-seq analysis using luminal epithelial specimens dissected out by laser capture microdissection revealed that the signaling pathways related to extracellular matrix, cell adhesion, and cell proliferation are altered in Pgr fl/flLtf Cre/+ mice. These findings suggest that epithelial PGR controls both epithelial and stromal P4 responsiveness and epithelial cell differentiation, which provides normal uterine receptivity and subsequent embryo attachment.

Clinicopathologic characteristics of epithelial ovarian tumours in Ile-Ife, Nigeria

Background: Epithelial ovarian tumours (EOT) have complex clinicopathologic characteristics and biological behaviours. There are benign, borderline and malignant ovarian tumours and the commonest ovarian tumours in many regions are of epithelial origin. Many studies have described the histomorphological characteristics of the tumours. Objective: To describe the clinical and histopathological features of epithelial ovarian tumours. Methods: This was a retrospective review of the histopathology reports of all epithelial ovarian tumours specimens submitted to the Department of Morbid Anatomy and Forensic Medicine, Obafemi Awolowo University Teaching Hospital, Ile-Ife from January 2005 to December 2014. The EOT cases were described in terms of age, clinicopathological characteristics and distribution of histological types. Results: The frequencies of benign, borderline and malignant EOTs were 41.2%, 3.9% and 54.9% respectively and the patients were aged 23 to 94 years (mean 46.5±2.6 years). The majority of cases were often asymptomatic. Conclusion: Abdominal swelling was the most common presenting complaint while serous ovarian tumours were the most preponderant histological types.

Nuclear Factor κB/MicroRNA-155 Upregulates the Expression Pattern of Cytokines in Regulating the Relapse of Chronic Sinusitis with Nasal Polyps and the Underlying Mechanism of Glucocorticoid.

BackgroundThe aim of this study was to explore the upregulated nuclear factor κB (NF-κB)/microRNA-155 (miR-155) in regulating inflammatory responses and relapse of chronic rhinosinusitis (CRS) with nasal polyps (NP), which underlies the molecular mechanism of glucocorticoid treatment.Material/MethodsThe study recruited 25 patients with eosinophilic (Eos) CRSwNP, 25 patients with Non-Eos CRSwNP, 25 patients with CRS without NP (CRSsNP) and 30 patients with nasal septum deviation (control group). The expression of NF-κB/miR-155 and inflammatory cytokines was detected in epithelial tissue specimens. Additionally, a mouse model of Eos CRSwNP was established, and the mice were treated by NF-κB inhibitor, miR-155 antagomir, or dexamethasone (DEX) to explore the role of NF-κB/miR-155 and the anti-inflammatory effects of glucocorticoid treatment.ResultsResults showed that the expression level of NF-κB/miR-155 was significantly elevated in the Eos CRSwNP group, accompanied by the upregulation of cytokines: tumor necrosis factor (TNF)-α, interleukin (IL)-1, IL-4, IL-5 (P<0.05) compared with the control group, the CRSsNP group or the Non-Eos CRSwNP group. The upregulation of NF-κB/miR-155 increased inflammatory mediator cyclooxygenase2 (COX2) while decreasing anti-inflammatory mediator Src homology-2 domain-containing inositol 5-phosphatase 1 (SOCS1), which resulted in the aberrant expression pattern of cytokines in the mice model. DEX treatment inhibited the expression of cytokines and decreased the relapse rate of Eos CRSwNP via inhibiting NF-κB/miR-155 (P<0.05).ConclusionsThe upregulation of NF-κB/miR-155 was crucial in mediating the aberrant expression of inflammatory cytokines in Eos CRSwNP. This molecular mechanism is a concern with the high relapse rate of Eos CRSwNP. However, glucocorticoid treatment inhibited the relapse of CRSwNP via downregulation of NF-κB/miR-155.