Healthy Tissue Samples Research Articles

Passive mechanical properties in healthy and infarcted rat left ventricle characterised via a mixture model

During the cardiac cycle, electrical excitation is coupled with mechanical response of the myocardium. Besides the active contraction, passive mechanics plays an important role, and its behaviour differs in healthy and diseased hearts as well as among different animal species. The aim of this study is the characterisation of passive mechanical properties in healthy and infarcted rat myocardium by means of mechanical testing and subsequent parameter fitting. Elasticity assessments via uniaxial extension tests are performed on healthy and infarcted tissue samples from left ventricular rat myocardium. In order to fully characterise the orthotropic cardiac tissue, our experimental data are combined with other previously published tests in rats – shear tests on healthy myocardium and equibiaxial tests on infarcted tissue. In a first step, we calibrate the Holzapfel-Ogden strain energy function in the healthy case. Sa far, this orthotropic constitutive law for the passive myocardium has been fitted to experimental data in several species, however there is a lack of an appropriate parameter set for the rat. With our determined parameters, a finite element simulation of the end-diastolic filling is performed. In a second step, we propose a model for the infarcted tissue. It is represented as a mixture of intact myocardium and a transversely isotropic scar structure. In our mechanical experiments, the tissue after myocardial infarction shows significantly stiffer behaviour than in the healthy case, and the stiffness correlates with the amount of fibrosis. A similar relationship is observed in the computational simulation of the end-diastolic filling. We conclude that our new proposed material model can capture the behaviour of two kinds of tissues – healthy and infarcted rat myocardium, and its calibration with the fitted parameters represents the experimental data well.

LncRNA TP73-AS1 enhances the malignant properties of pancreatic ductal adenocarcinoma by increasing MMP14 expression through miRNA -200a sponging.

Pancreatic ductal adenocarcinoma (PDAC) is an invasive and aggressive cancer that remains a major threat to human health across the globe. Despite advances in cancer treatments and diagnosis, the prognosis of PDAC patients remains poor. New and more effective PDAC therapies are therefore urgently required. In this study, we identified a novel host factor, namely the LncRNA TP73‐AS1, as overexpressed in PDAC tissues compared to adjacent healthy tissue samples. The overexpression of TP‐73‐AS1 was found to correlate with both PDAC stage and lymph node metastasis. To reveal its role in PDCA, we targeted TP73‐AS1 using LnRNA inhibitors in a range of pancreatic cancer (PC) cell lines. We found that the inhibition of TP73‐AS1 led to a loss of MMP14 expression in PC cells and significantly inhibited their migratory and invasive capacity. No effects of TP73‐AS1 on cell survival or proliferation were observed. Mechanistically, we found that TP73‐AS1 suppressed the expression of the known oncogenic miR‐200a. Taken together, these data highlight the prognostic potential of TP73‐AS1 for PC patients and highlight it as a potential anti‐PDAC therapeutic target.

Comparison of the Allelic Alterations between InDel and STR Markers in Tumoral Tissues Used for Forensic Purposes

Background and objectives: Over the last two decades, human DNA identification and kinship tests have been conducted mainly through the analysis of short tandem repeats (STRs). However, other types of markers, such as insertion/deletion polymorphisms (InDels), may be required when DNA is highly degraded. In forensic genetics, tumor samples may sometimes be used in some cases of human DNA identification and in paternity tests. Nevertheless, tumor genomic instability related to forensic DNA markers should be considered in forensic analyses since it can compromise genotype attribution. Therefore, it is useful to know what impact tumor transformation may have on the forensic interpretation of the results obtained from the analysis of these polymorphisms. Materials and Methods: The aim of this study was to investigate the genomic instability of InDels and STRs through the analysis of 55 markers in healthy tissue and tumor samples (hepatic, gastric, breast, and colorectal cancer) in 66 patients. The evaluation of genomic instability was performed comparing InDel and STR genotypes of tumor samples with those of their healthy counterparts. Results: With regard to STRs, colorectal cancer was found to be the tumor type affected by the highest number of mutations, whereas in the case of InDels the amount of genetic mutations turned out to be independent of the tumor type. However, the phenomena of genomic instability, such as loss of heterozygosity (LOH) and microsatellite instability (MSI), seem to affect InDels more than STRs hampering genotype attribution. Conclusion: We suggest that the use of STRs rather than InDels could be more suitable in forensic genotyping analyses given that InDels seem to be more affected than STRs by mutation events capable of compromising genotype attribution.

La asociación entre Fusobacterium nucleatum y el cáncer colorrectal: una revisión sistemática y metaanálisis

IntroductionThe etiological factors of colorectal cancer (CRC) are not precisely known, although genetic and environmental factors have been implicated. A possible association with Fusobacterium nucleatum may provide opportunities for an early diagnosis. ObjectiveTo review studies that address the association between F. nucleatum and CRC. MethodsThe MEDLINE PubMed database was searched using the terms «colorectal cancer» and «Fusobacterium nucleatum», retrieving publications published up to January 1 2020. Stata software was used for a meta-analysis. ResultsThe systematic review included 57 articles. Meta-analysis results indicated a more frequent presence of F. nucleatum in CRC tumor tissue samples in comparison to control samples of healthy tissue, with an odds ratio of 4.558 (95% CI: 3.312-6.272), and in comparison, to control samples of colorectal adenomas, with an odds ratio of 3.244 (95% CI: 2.359-4.462). ConclusionThere is a more frequent presence of F. nucleatum in the CRC. However, further studies are needed to verify this relationship.

Transcriptional Biomarkers in Oral Cancer: An Integrative Analysis and the Cancer Genome Atlas Validation.

Objective:An impervious mortality rate in oral cancer (OC) to a certain extent explains the exigencies of precise biomarkers. Therefore, the study was intended to identify OC candidate biomarkers using samples of healthy normal tissues (N=335), adjacent normal tissues (N=93) and OC tissues (N=533) from online microarray data. Methods:Differentially expressed genes (DEGs) were recognised through GeneSpring software (Fold change >4.0 and ‘p’ value <0.001 with Benjamini Hochberg false discovery rate). The DEGs were analysed for their functional annotation and network using GeneCodis 4.0 and STRING databases. The DEGs were further cross-examined in the cancer genome atlas (TCGA) database. Results:197, 229 and 104 DEGs were spotted from three comparisons: i) OC tissues against healthy normal tissues, ii) OC tissues against adjacent normal tissues and iii) adjacent normal tissues against healthy normal tissues, respectively. Functional gene set enrichment unveiled significance of focal adhesion pathway in OC initiation and progression. Advanced analysis of TCGA cohort (n=345) recognised 85 genes that were altered in ≥10% OC patients for mutations, copy number variations, m-RNA expression and protein expression. Strikingly, the elected 5-gene panel (YWHAZ, RHOA, DLG1, LY6E and PLEC) showed the location on chromosome 3 and 8 and each gene was found to be altered in ≥20% OC patients for m-RNA expression. Further, TCGA validation demonstrated significant association of EIF4A2, CTNNA1 and PMEPA1 expression with overall and disease-free survival. Conclusion: Using this integrative approach, our study identified prominent transcriptional biomarkers which may be significant targets for OC therapy. Additional validation of these biomarkers in experimental prospective and retrospective studies will launch them in OC clinics.

CD8 and CD4 T Cell Populations in Human Kidneys.

Background: At border sites, and in internal organs, tissue resident memory T cells (TRM) contribute to the immune barrier against pathogens like viruses, bacteria, fungi, and cancer. However, information on the presence and function of these cells in the human kidney is scant. In order to better understand the T cell-mediated immunological defense in this organ, we aimed to determine phenotypic and functional aspects of CD8 and CD4 T cells present in healthy and allograft kidney tissue. Methods: Using multichannel flow cytometry, we assessed the phenotype and function of T cells in healthy renal tissue samples (n = 5) and kidney allograft tissue (n = 7) and compared these aspects to T cells in peripheral blood from healthy controls (n = 13). Results: Kidney tissue samples contained substantial amounts of CD8 and CD4 T cells. In contrast to the circulating cells, kidney T cells frequently expressed CD69 and CD103, and were more often actively cycling. Furthermore, nearly all kidney T cells expressed CXCR3, and often expressed CXCR6 compared to T cells in the circulation. Markedly, kidney T cells produced greater quantities of IFNγ than circulating cells and were frequently polyfunctional. Conclusion: Functional T cells with the characteristic traits of TRM reside in human kidney tissues. These cells are more often actively cycling and frequently express CXCR3 and CXCR6.

Novel Identified Circular Transcript of RCAN2, circ-RCAN2, Shows Deviated Expression Pattern in Pig Reperfused Infarcted Myocardium and Hypoxic Porcine Cardiac Progenitor Cells In Vitro.

Circular RNAs (circRNAs) are crucial in gene regulatory networks and disease development, yet circRNA expression in myocardial infarction (MI) is poorly understood. Here, we harvested myocardium samples from domestic pigs 3 days after closed-chest reperfused MI or sham surgery. Cardiac circRNAs were identified by RNA-sequencing of rRNA-depleted RNA from infarcted and healthy myocardium tissue samples. Bioinformatics analysis was performed using the CIRIfull and KNIFE algorithms, and circRNAs identified with both algorithms were subjected to differential expression (DE) analysis and validation by qPCR. Circ-RCAN2 and circ-C12orf29 expressions were significantly downregulated in infarcted tissue compared to healthy pig heart. Sanger sequencing was performed to identify the backsplice junctions of circular transcripts. Finally, we compared the expressions of circ-C12orf29 and circ-RCAN2 between porcine cardiac progenitor cells (pCPCs) that were incubated in a hypoxia chamber for different time periods versus normoxic pCPCs. Circ-C12orf29 did not show significant DE in vitro, whereas circ-RCAN2 exhibited significant ischemia-time-dependent upregulation in hypoxic pCPCs. Overall, our results revealed novel cardiac circRNAs with DE patterns in pCPCs, and in infarcted and healthy myocardium. Circ-RCAN2 exhibited differential regulation by myocardial infarction in vivo and by hypoxia in vitro. These results will improve our understanding of circRNA regulation during acute MI.

Gene expression of one important microRNA in blood and tissue samples of oral squamous cell carcinoma

Introduction: Oral Squamous Cell Carcinoma (OSCC) is one of the most common oral malignancies, which accounts for 80-90% of malignant neoplasms of the oral cavity. MicroRNAs (miRNAs) are small RNA molecules that regulate post-transcriptional gene expression by targeting mRNAs. Materials and Methods: In this case-control study, 40 patients with oral squamous cell carcinoma and 40 healthy individuals as control were studied. Blood samples were collected from both groups. Also, 30 cancer tissue samples and 30 healthy tissue samples were prepared and evaluated. RNA was extracted from collected peripheral blood and tissue samples and evaluated for the expression level of miR-494 via real-time PCR technique. P. value values&lt;0.05 were considered statistically significant. Results: The expression level of miR-494 in serum (peripheral blood) of patients with oral squa- mous cell carcinoma increased by 1.12 fold (P-value&lt;0.001) compared with healthy individuals. Also, the expression level of miR-494 in samples of oral squamous cell carcinoma infected tissue showed a 1.28-fold increase compared to healthy tissue. Conclusion: The results of this study indicate an increase in the expression level (up-regula- tion) of miR-494 in oral squamous cell carcinoma. This biomarker can be used in screening and early detection of oral squamous cell carcinoma.

MYBPC2 and MYL1 as Significant Gene Markers for Rhabdomyosarcoma.

Background:Rhabdomyosarcoma is the most common soft tissue tumor in children. Rhabdomyosarcoma commonly results in pain and bleeding caused by tumor compression and is prone to early metastasis and recurrence, which can seriously affect the therapeutic outcomes and long-term prognosis. Up to 37.7% of rhabdomyosarcomas may metastasize. Therefore, the molecular mechanisms underlying rhabdomyosarcoma must be explored to identify an effective target for its early diagnosis and specific treatment.Methods:A dataset of 18 rhabdomyosarcoma tissue samples and 6 healthy skeletal muscle samples was downloaded. Differentially expressed genes between rhabdomyosarcoma and healthy tissue samples were identified by GEO2R. Kyoto Encyclopedia of Genes and Genomes and gene ontology pathway enrichment analyses were performed. A protein–protein interaction network was constructed, and hub genes were identified. Expression and survival analyses of hub genes were performed. Additionally, 30 patients with rhabdomyosarcoma were recruited, and overall survival information and samples were collected. Reverse transcription quantitative real-time polymerase chain reaction assays were performed to verify the expression of MYBPC2 and MYL1 in rhabdomyosarcoma tumor tissues. The Kaplan–Meier method was used to explore overall survival based on our clinical data.Results:In total, 164 genes were up-regulated and 394 were down-regulated in rhabdomyosarcoma tumor tissues. Gene ontology analysis revealed that variations were predominantly enriched in the cell cycle, muscle contraction, muscle system processes, cytoskeleton, nucleotide binding, and cytoskeletal protein binding. The protein–protein interaction network revealed 3274 edges, and 441 nodes were constructed. Ten hub genes were identified; of these, MYBPC2 and MYL1 were significantly up-regulated in rhabdomyosarcoma. Compared with the healthy group, patients with rhabdomyosarcoma exhibiting high expression of MYBPC2 and MYL1 exhibited significantly worse overall survival.Conclusions:We found differentially expressed genes between rhabdomyosarcoma and healthy tissue samples. MYBPC2 and MYL1 may be involved in the pathogenesis of rhabdomyosarcoma and therefore deserve further exploration.

Активирующие мутации генов PI3K/AKT/mTOR- сигнального пути и состояние овариального резерва у пациенток репродуктивного возраста с глубоким инфильтративным эндометриозом

Objective. To study the effect of deep infiltrating endometriosis on the status of ovarian reserve in patients of reproductive age and to assess the spectrum of PIK3CA gene mutations among patients with infiltrating form of external genital endometriosis. Patients and methods. The study group included 50 patients of reproductive age with deep infiltrating endometriosis, of whom 18 had deep infiltrating endometriosis combined with ovarian endometriomas. The comparison group included 25 patients of reproductive age who underwent laparoscopic metroplasty of post-cesarean section uterine scar. In all patients, serum levels of anti-mullerian hormone (AMH), follicle-stimulating hormone, and estradiol were determined by enzyme immunoassay, and an antral follicle count in the ovaries was done via transvaginal ultrasound. The search for activating mutations in the PIK3CA gene was performed by next-generation DNA sequencing in the samples of ovarian endometriomas from patients with a combination of infiltrating endometriosis and endometrioid cysts (n = 18) and in biopsy samples of healthy ovarian tissue from all patients in the study (n = 50) and comparison groups (n = 25). Results. Evaluation of the ovarian reserve status in patients from two groups showed that levels of AMH were lower in patients with infiltrating form of external genital endometriosis than in the comparison group by 1.0 ng/mL on average (2.6 ± 2.2 ng/mL in the study group, 3.6 ± 3.5 ng/mL in the comparison group), but the difference was not statistically significant, p &gt; 0.05. The number of antral follicles according to transvaginal ultrasound was significantly lower in the study group (8.5 ± 4.5) than in the comparison group (12.2 ± 4.1), p = 0.001. This difference was statistically significant both for patients with ovarian endometriomas (6.0 ± 4.2, p &lt; 0.001) and for patients without ovarian endometrioid tumors (9.8 ± 4.2, p = 0.04). Our study did not detect PIK3CA gene mutations in any of the ovarian endometrioma tissue samples from patients with a combination of infiltrating endometriosis and endometrioid cysts, and in none of the healthy ovarian tissue biopsy samples from patients in the study and comparison groups. Conclusion. Thus, the presence of deep infiltrating endometriosis is associated with diminished ovarian reserve in patients of reproductive age, regardless of the presence of ovarian endometrioid tumors. Population studies are needed to detect PIK3CA gene mutations in endometriosis, as well as to investigate mutations in other genes encoding regulatory proteins of the PI3K/AKT/mTOR anti-apoptotic signaling pathway to reveal the mechanisms of ovarian reserve depletion in case of infiltrating forms of external genital endometriosis. Key words: infertility, deep infiltrating endometriosis, ovarian reserve, PI3K/AKT/mTOR signaling pathway, ovarian endometrioma

The role of adiponectin and its receptor in patients with idiopathic membranous nephropathy complicated with hyperuricemia

Introduction/Objective. This study aimed to assess the changes of adiponectin (APN), IL-1?, adiponectin receptor 1 (Adipo R1), and NLRP3 expression of patients with idiopathic membranous nephropathy (IMN) complicated with hyperuricemia (HUA) and analyze the relationship between the APN pathway and the NLRP3 pathway. Methods. A group of 48 patients with IMN + HUA, a group of 49 patients with IMN, 30 healthy controls, and 24 samples of healthy renal tissue were evaluated. APN and IL-1? of each group were detected by the ELISA method. AdipoR1 and NLRP3 in kidney tissue were detected by immunohistochemistry. The clinical data of each group were collected, and the relationship between APN, IL-1?, AdipoR1, NLRP3, and other indexes was analyzed. Results. (1) The concentration of UA, APN, IL-1?, and NLRP3 in the IMN + HUA group were significantly higher than those in the IMN group, but the AdipoR1 was lower. (2) With the severity of chronic kidney disease stage, APN, IL-1?, and NLRP3 gradually increased in the IMN + HUA group, but AdipoR1 gradually decreased. However, the aforementioned indicators did not change significantly in the IMN stages. Conclusion. The AdipoR1?AMPK and NLRP3?caspase-1?IL-1? signaling pathway may play an essential role in IMN + HUA patients. An intervention on these two pathways may have significant impact on the disease occurrence and progression in IMN + HUA patients.

Evaluation of TGF-β1 and EGFR in Cleft Affected Lip Mucosa.

Background.The morphopathogenesis of orofacial cleft development is only partly understood; therefore, it is important to identify factors, which possibly could be involved in it. The aim of the study was to evaluate the distribution of TGF-β1 and EGFR-containing cells in cleft affected lip mucosa.Materials and Methods.The study group included lip mucosa tissue samples from 14 patients with orofacial cleft. The control group contained 11 healthy oral mucosa tissue samples. The tissue sections were stained by immunohistochemistry for TGF-β1 and EGFR. The expression of positive structures was graded semiquantitatively. IBM SPSS 26.0 was used for statistical analysis, Spearman`s rank correlation and Mann-Whitney U tests were performed.Results.Mostly few to moderate number (+/++) of TGF-β1-containing cells was found in epithelium, also the same number of fibroblasts and macrophages was seen in the lamina propria of cleft affected lip mucosa. Meanwhile, healthy oral mucosa on average demonstrated a moderate number (++) of TGF-β1-containing epithelial cells, fibroblasts, and macrophages. A variable, mostly indistinct number of EGFR-containing cells was seen in the epithelium of cleft affected lip mucosa, meanwhile, mostly no (0) EGFR positive cells were found in the epithelium of healthy mucosa. Statistically significantly less TGF-β1-containing cells were found in the epithelium of cleft affected lip mucosa than in the healthy mucosa (U=33.000; p=0.015). Also, the lamina propria of cleft affected lip mucosa showed statistically significantly less TGF-β1 immunoreactive fibroblasts and macrophages than the healthy mucosa (U=28.500; p=0.006).Conclusions.The decreased number of TGF-β1-containing epithelial cells, fibroblasts and macrophages in cleft affected lip mucosa proves the role of problematic tissue remodelation in the cleft pathogenesis. The distribution of EGFR in cleft affected and healthy mucosa is similar and possibly does not play a role in the cleft development of humans.

The role of long non-coding RNA (lncRNA) in the development of ovarian cancer

The aim of this study was to review research on the role of long non-coding RNA (lncRNA) in ovarian cancer. This article analyses studies on the effect of increased lncRNA expression on the size of ovarian cancer and the incidence of metastasis. The review covers a period from October 15, 2018 to August 22, 2020, and comprises 23 studies in which a total of 1,580 women with ovarian cancer participated, and an undetermined number of control groups where healthy tissue samples were collected. A review of the studies indicates that increased lncRNA expression is associated with elevated ovarian cancer size and metastatic risk. The most studied lncRNA include HOTAIR, CCAT2, GAS5, MALAT-1, UCA1. Studies assessing the expression levels of HOTAIR lncRNA and CCAT2 in normal and cancer tissue showed varying levels of expression in studies of different authors, which indicates that the expression of the same lncRNA may vary individually or is a result of study errors.

Investigating Serum and Tissue Expression Identified a Cytokine/Chemokine Signature as a Highly Effective Melanoma Marker.

Simple SummaryIn this study, we investigated the expression of 27 cytokines/chemokines in the serum of 232 individuals (136 melanoma patients vs. 96 controls). It identified several cytokines/chemokines differently expressed in melanoma patients as compared to the healthy controls, as a function of the presence of the melanoma, age, tumor thickness, and gender, indicating different systemic responses to the melanoma presence. We also analyzed the gene expression of the same 27 molecules at the tissue level in 511 individuals (melanoma patients vs. controls). From the gene expression analysis, we identified several cytokines/chemokines showing strongly different expression in melanoma as compared to the controls, and the 4-gene signature “IL-1Ra, IL-7, MIP-1a, and MIP-1b” as the best combination to discriminate melanoma samples from the controls, with an extremely high accuracy (AUC = 0.98). These data indicate the molecular mechanisms underlying melanoma setup and the relevant markers potentially useful to help the diagnosis of biopsy samples.The identification of reliable and quantitative melanoma biomarkers may help an early diagnosis and may directly affect melanoma mortality and morbidity. The aim of the present study was to identify effective biomarkers by investigating the expression of 27 cytokines/chemokines in melanoma compared to healthy controls, both in serum and in tissue samples. Serum samples were from 232 patients recruited at the IDI-IRCCS hospital. Expression was quantified by xMAP technology, on 27 cytokines/chemokines, compared to the control sera. RNA expression data of the same 27 molecules were obtained from 511 melanoma- and healthy-tissue samples, from the GENT2 database. Statistical analysis involved a 3-step approach: analysis of the single-molecules by Mann–Whitney analysis; analysis of paired-molecules by Pearson correlation; and profile analysis by the machine learning algorithm Support Vector Machine (SVM). Single-molecule analysis of serum expression identified IL-1b, IL-6, IP-10, PDGF-BB, and RANTES differently expressed in melanoma (p < 0.05). Expression of IL-8, GM-CSF, MCP-1, and TNF-α was found to be significantly correlated with Breslow thickness. Eotaxin and MCP-1 were found differentially expressed in male vs. female patients. Tissue expression analysis identified very effective marker/predictor genes, namely, IL-1Ra, IL-7, MIP-1a, and MIP-1b, with individual AUC values of 0.88, 0.86, 0.93, 0.87, respectively. SVM analysis of the tissue expression data identified the combination of these four molecules as the most effective signature to discriminate melanoma patients (AUC = 0.98). Validation, using the GEPIA2 database on an additional 1019 independent samples, fully confirmed these observations. The present study demonstrates, for the first time, that the IL-1Ra, IL-7, MIP-1a, and MIP-1b gene signature discriminates melanoma from control tissues with extremely high efficacy. We therefore propose this 4-molecule combination as an effective melanoma marker.

Plasma levels and tissue expression of soluble TGF\u03b2rIII receptor in women with early-stage breast cancer and in healthy women: a prospective observational study

BackgroundMammary carcinogenesis is partly regulated by the transforming growth factor beta (TGFβ) signaling pathway. Its function in cancer progression and metastasis is highly dependent on disease stage, and it is likely modulated by the ratio of membrane-bound vs. soluble TGFβrIII (sTGFβrIII). In this prospective observational study, we assessed tissue expression and plasma levels of sTGFβrIII in healthy women, women with benign breast lesions and in early-stage breast cancer patients.MethodsIn a preliminary study, plasma sTGFβrIII levels were determined in 13 healthy women (age 19–40 years) at different phases of the ovarian cycle, and in 15 patients (age 35–75 years) at different times of the day. The main study assessed plasma concentrations of sTGFβrIII in: (i) 158 healthy women in whom breast lesions were excluded; (ii) 65 women with benign breast lesions; (iii) 147 women with newly diagnosed breast cancer classified as American Joint Committee on Cancer (AJCC) stages 0 to IIB. Completers provided blood samples before surgery and at 10–30 and 160–180 days after surgery. Plasma sTGFβrIII concentrations were determined using an indirect ELISA kit. Part of the removed tissues underwent immunohistochemical (IHC) staining and analysis of tissue TGFβrIII expression.ResultsThere appeared no relevant variations in plasma sTGFßrIII levels at different times of the day or different ovarian cycle phases. Before surgery, breast cancer patients had somewhat higher sTGFβrIII than healthy women, or those with benign breast lesions (by 14.5 and 26 ng/mL, respectively), with a tendency of larger differences at higher age. This correlated with lower expression of TGFβrIII in breast cancer vs. healthy tissue samples. At 160–180 days after surgery, plasma sTGFβrIII levels in breast cancer patients declined by 23–26 ng/mL.ConclusionsPlasma sTGFβrIII levels do not seem to relevantly vary during the day or the ovarian cycle. The coinciding higher plasma levels in newly diagnosed cancer patients than in healthy subjects and lower TGFβrIII expression in the malignant than in healthy breast tissue suggest ectodomain shedding as a source of circulating sTGFβrIII. Decline in plasma levels after tumor removal supports such a view.

Effects of cold physical plasma on oral lichen planus: An in vitro study (Effects of CAP on OLP).

In the search for more effective and safe treatment avenues, we investigated cold physical plasma as a new treatment modality for therapy of oral lichen planus (OLP). Healthy and diseased human mucosal tissue samples with a size of 3mm in diameter obtained from OLP patients were subjected to plasma treatment ex vivo or were left untreated. Tissue sections were quantified for immune-infiltration of CD4+ , CD8+ , CD45RA+ , and CD45R0+ T cells. Moreover, the tissues' inflammatory profile was assessed by analyzing 12 different cytokines in the surrounding media. A significantly increased infiltrate of CD8+ and CD45-R0+ T cells was detected in OLP tissue samples when compared to healthy tissue. A higher concentration of interleukin (IL) 1β, IL6, IL8, and granulocyte macrophage-colony stimulating factor (GM-CMF) was detected in OLP samples compared to healthy mucosal tissue. For all cytokines and chemokines investigated, 23 out of 24 comparisons showed a decrease in tendency (significant for IL1β, IL2, IL10, and GM-CSF) in response to plasma treatment. In ex vivo-treated tissue, a decrease of T-cell infiltrate in OLP lesions compared with healthy tissue was observed. Our findings suggest cold physical plasma can be a promising therapeutic option for OLP that requires further validation in vivo.

Regulation of ABCG2 expression by Wnt5a through FZD7 in human pancreatic cancer cells.

ATP-binding cassette subfamily G member 2 (ABCG2), a member of the ABC transporter superfamily, has been implicated in the development of chemotherapeutic drug resistance in cancer cells. However, the regulators of ABCG2 expression and their roles in anticancer drug resistance have not been fully characterized, especially in the context of pancreatic cancer. The aim of the present study was to investigate whether ABCG2 contributed to drug resistance in pancreatic cancer and to elucidate its regulatory molecular pathways. Using immunohistochemical analysis of pancreatic ductal adenocarcinoma and adjacent healthy tissue samples, the present study identified a positive correlation between ABCG2 and Wnt5a, a member of the Wnt family of secreted proteins. It was also determined that treatment with recombinant human Wnt5a protein could upregulate the expression of ABCG2 in the Capan-2 human pancreatic cancer cell line and enhance its resistance to gemcitabine. The upregulation of ABCG2 by Wnt5a was inhibited by small interfering RNA silencing of Frizzled class receptor 7 (FZD7) or by FZD7 inhibitors. Moreover, both FZD7 silencing or inhibition of its function attenuated gemcitabine resistance induced by Wnt5a in Capan-2 cells. Therefore, the present findings suggested that Wnt5a and FZD7 acted as upstream regulators of ABCG2 expression and that FZD7 may be an essential factor for Wnt5a-induced gemcitabine resistance in pancreatic cancer cells.

Serum and tissue expression levels of microRNAs-661, -571 and -770-5p among diabetic foot ulcer patients compared to healthy controls

BackgroundMicro-RNAs (miRNAs) are considered as biomarkers playing a role for prediction of type 2 diabetes mellitus (T2DM) and diabetic foot ulcer (DFU). More relevantly, miRNAs confer higher sensitivity levels than currently used tools. In this study, we aimed to evaluate serum and tissue expression levels of miR-770-5p, miR-571 and miR-661 among T2DM and DFU patients compared to healthy control population. MethodsThe study included 600 subjects (300 patients, 150 healthy control individuals and 150 individuals with trauma injury). Sampling was taken from T2DM (blood) and DFU patients (blood and tissue), healthy subjects (blood control samples) and individuals with trauma injury (tissue control samples). Serum and tissue expression levels of miR-770-5p, miR-571 and miR-661 were measured using quantitative Real time PCR (RT-qPCR) technique. ResultsThe miR-661 expression was significantly higher in tissue and sera samples of patients with DFU compared to control groups (p < 0.001 and p = 0.003), but not from sera of T2DM patients (without DFU). In addition, the expression of miR-770 was significantly higher in diabetic tissue compared to healthy tissue (p = 0.002), healthy blood (p < 0.001) and sera of patients without DFU (0.0261). However, the miR-571 was not significantly different between T2DM patients' blood samples and each of healthy blood (p = 0.0134), healthy tissue (p = 0.1153) and other samples. Moreover, this miRNA expression level was not significantly different between sera of T2DM patients and healthy individuals (p = 0.142). ConclusionThe miR-661 and miR-770-5p had substantial roles as biomarkers in the rapid diagnosis of both T2DM and DFU diseases. Nonetheless, the miR-571 played no significant role as a diagnostic biomarker to this aim. Further in-depth studies are needed to verify the association of these miRNAs or other those potential miRNAs with the T2DM and DFU development.

The inhibitory effect of the deubiquitinase cylindromatosis (CYLD) on inflammatory responses in human gingival fibroblasts.

Experiments were performed to evaluate CYLD expression in human gingival tissue samples and to examine the effects of CYLD on inflammatory responses in lipopolysaccharide (LPS)- or TNF-α-stimulated human gingival fibroblasts (HGFs). Immunohistochemistry for CYLD and p65 expression was performed with healthy and inflamed gingival tissue samples. siRNA was used to knock down the expression of CYLD in HGFs. Upon LPS or TNF-α stimulation, NF-κB activation was detected in control and CYLD-knockdown HGFs. RT-PCR was applied to determine gene expression. Western blot analyses were employed to assess protein expression. Immunofluorescence staining was carried out to evaluate the nuclear translocation of p65. Immunohistochemical staining showed the expression of CYLD in human gingival tissues. In addition, CYLD protein expression was reduced in inflamed gingival tissue samples compared with healthy tissue samples. CYLD knockdown greatly enhanced the mRNA expression of proinflammatory cytokines in LPS- or TNF-α-stimulated HGFs. Furthermore, knocking down CYLD expression increased LPS-stimulated NF-κB activation in HGFs. Unexpectedly, CYLD knockdown did not affect TNF-α-induced NF-κB activation. Our results suggest that CYLD participates in periodontal inflammatory responses by negatively regulating LPS-induced NF-κB signalling.

Matrix metalloproteinase MMP1 and MMP9 genes expression in breast cancer tissue

Nowadays, Breast cancer is recognized as the most common cancer among women worldwide. Matrix metalloproteinases (MMPs) are a group of endopeptidase enzymes that can degrade extracellular matrix proteins and increase invasion and metastasis. The purpose of this study was to evaluate the expression of MMP1 and MMP9 genes in breast cancer tissue. In the present case-control study, 90 breast tissue sample, includes 45 breast cancer tissue samples as case group and 45 healthy breast tissue samples as control group were studied. After RNA extraction and cDNA synthesis, MMP1 and MMP9 genes mRNA expression levels were evaluated using Real-Time PCR. In the present study, a significant increase was observed in the expression levels of MMP1 and MMP9 genes mRNA expression in breast cancer tissues compared to normal peripheral tissues. The mRNA expression levels of MMP1 and MMP9 genes in breast cancer tissue were increased by 2.5 and 2.8 fold, respectively. Due to the increased expression of MMP1 and MMP9 genes in breast cancer tissue samples, these proteins can be used as diagnostic markers and novel therapeutic targets. However, further studies with larger sample sizes and on other MMPs are recommended.