Background Diabetic foot ulceration is a serious chronic complication of diabetes mellitus characterized by high disability, mortality, and morbidity. Platelet-rich plasma (PRP) has been widely used for diabetic wound healing due to its high content of growth factors. However, its application is limited due to the rapid degradation of growth factors. The present study aimed to evaluate the efficacy of combined adipose-derived mesenchymal stem cells (ADSCs) and PRP therapy in promoting diabetic wound healing in relation to the Notch signaling pathway. Methods Albino rats were allocated into 6 groups [control (unwounded), sham (wounded but non-diabetic), diabetic, PRP-treated, ADSC-treated, and PRP+ADSCs-treated groups]. The effect of individual and combined therapy was evaluated by assessing wound closure rate, epidermal thickness, dermal collagen, and angiogenesis. Moreover, gene and protein expression of key elements of the Notch signaling pathway (Notch1, Delta-like canonical Notch ligand 4 (DLL4), Hairy Enhancer of Split-1 (Hes1), Hey1, Jagged-1), gene expression of angiogenic marker (vascular endothelial growth factor and stromal cell-derived factor 1) and epidermal stem cells (EPSCs) related gene (ß1 Integrin) were assessed. Results Our data showed better wound healing of PRP+ADSCs compared to their individual use after 7 and 14 days as the combined therapy caused reepithelialization and granulation tissue formation with a marked increase in area percentage of collagen, epidermal thickness, and angiogenesis. Moreover, Notch signaling was significantly downregulated, and EPSC proliferation and recruitment were enhanced compared to other treated groups and diabetic groups. Conclusions These data demonstrated that PRP and ADSCs combined therapy significantly accelerated healing of diabetic wounds induced experimentally in rats via modulating the Notch pathway, promoting angiogenesis and EPSC proliferation.
Background: Neonatal sepsis is considered as a complicated syndrome, which requires urgent intervention to avoid the unfavorable outcome. Thus, biomarkers that can either distinguish sepsis early or predict sepsis outcome are of critical need. Therefore, the aim of the current study was to investigate the clinical value of miR-187, miR-101, and miR-21 on neonatal sepsis diagnosis and prediction of prognosis. Fifty neonates with sepsis, 30 neonates with SIRS, and 20 healthy neonates were selected. Relative expression levels of the selected miRNAs were quantified by qRT-PCR. Serum CRP and PCT were analyzed. Results: miR-101 and miR-187 expression levels were elevated in septic neonates compared with SIRS neonates and normal controls. The AUC of miR-101, miR-187, and PCT to predict sepsis diagnosis were 0.908, 789, and 0.856, respectively. miR-21 expression levels in non-survivors were significantly higher than in survivors. The AUC of miR-21, a score of neonatal acute physiology (SNAP-II), and PCT to detect the predictive mortality value were 0.793, 0.781, and 0.635, respectively. Survival analysis revealed that high miR-21 expression levels were related to low survival rates. miR-21 and SNAP II were independent risk factors for sepsis mortality, and the AUC of the two combined variables' predictive probabilities was 0.926 and yielded a specificity of 91.2% and a sensitivity of 81.3%, which was higher than that of either miR-21 or SNAP II. Conclusion: miR-101 might function as a hopeful diagnostic biomarker for neonatal sepsis. Additionally, miR-21 gained attention to be a valuable predictor for sepsis prognosis especially if combined with SNAP II.
Background Diabetic foot ulceration is a serious chronic complication of diabetes mellitus characterized by high disability, mortality and morbidity. Platelet-rich plasma (PRP) has been widely used for diabetic wound healing due to its high content of growth factors. However, its application is limited due to rapid degradation of growth factors. The present study aimed to evaluate the efficacy of combined adipose derived mesenchymal stem cells (ADSCs) and PRP therapy in promoting diabetic wound healing in relation to the Notch signaling pathway. Methods Albino rats were allocated into 6 groups (control, sham, diabetic, PRP-treated, ADSCs-treated and PRP+ADSCs-treated groups). The effect of individual and combined therapy was evaluated by assessing wound closure rate, epidermal thickness, dermal collagen and angiogenesis. Moreover, gene and protein expression of key elements of Notch signaling pathway (Notch1, Delta like canonical Notch ligand 4 (DLL4), Hairy Enhancer of Split-1 (Hes1), Hey1, Jagged-1), gene expression of angiogenic marker (Vascular endothelial growth factor & stromal cell-derived factor 1) and epidermal stem cells (EPSCs) related gene (ß1 Integrin) were assessed. Results Our data showed a strong wound-healing effect of PRP+ADSCs compared to their individual use after 7 and 14 days. Combined therapy caused marked increase in area percentage of collagen, epidermal thickness and angiogenesis. Moreover, Notch signaling was significantly down-regulated, EPSCs proliferation and recruitment was enhanced compared to other treated groups and diabetic group. Conclusions These data demonstrated that PRP and ADSCs combined therapy significantly accelerated healing of diabetic wounds induced experimentally in rats via modulating Notch pathway, promoting angiogenesis and EPSCs proliferation.
Background. Liver transplantation remains the only viable therapy for liver failure but has a severely restricted utility. Here, we aimed to decellularize rat livers to form acellular 3D bio-scaffolds suitable for seeding with induced pluripotent cells (iPSCs) as a tool to investigate the role of Wnt/β-catenin signaling in liver development and generation. Methods. Dissected rat livers were randomly divided into three groups: I (control); II (decellularized scaffolds) and III (recellularized scaffolds). Liver decellularization was established via an adapted perfusion procedure and assessed through the measurement of extracellular matrix (ECM) proteins and DNA content. Liver recellularization was assessed through histological examination and measurement of transcript levels of Wnt/β-catenin pathway, hepatogenesis, liver-specific microRNAs and growth factors essential for liver development. Adult rat liver decellularization was confirmed by the maintenance of ECM proteins and persistence of growth factors essential for liver regeneration. Results. iPSCs seeded rat decellularized livers displayed upregulated transcript expression of Wnt/β-catenin pathway-related, growth factors, and liver specification genes. Further, recellularized livers displayed restored liver-specific functions including albumin secretion and urea synthesis. Conclusion. This establishes proof-of-principle for the generation of three-dimensional liver organ scaffolds as grafts and functional re-establishment.
Background Very small embryonic-like stem cells (VSELs) are a rare population within the ovarian epithelial surface. They contribute to postnatal oogenesis as they have the ability to generate immature oocytes and resist the chemotherapy. These cells express markers of pluripotent embryonic and primordial germ cells. Objective We aimed to explore the capability of VSELs in restoring the postnatal oogenesis of chemo-ablated rat ovaries treated with bone marrow-derived mesenchymal stem cells (BM-MSCs) combined with pregnant mare serum gonadotropin (PMSG). Methods Female albino rats were randomly assigned across five groups: I (control), II (chemo-ablation), III (chemo-ablation + PMSG), IV (chemo-ablation + MSCs), and V (chemo-ablation + PMSG + MSCs). Postnatal oogenesis was assessed through measurement of OCT4, OCT4A, Scp3, Mvh, Nobox, Dazl4, Nanog, Sca-1, FSHr, STRA8, Bax, miR143, and miR376a transcript levels using qRT-PCR. Expression of selected key proteins were established as further confirmation of transcript expression changes. Histopathological examination and ovarian hormonal assessment were determined. Results Group V displayed significant upregulation of all measured genes when compared with group II, III or IV. Protein expression confirmed the changes in transcript levels as group V displayed the highest average density in all targeted proteins. These results were confirmed histologically by the presence of cuboidal germinal epithelium, numerous primordial, unilaminar, and mature Graafian follicles in group V. Conclusion VSELs can restore the postnatal oogenesis in chemo-ablated ovaries treated by BM-MSCs combined with PMSG.
Background: Previous reports have suggested the significant association of t(14;18) (q32;q21) and follicular lymphoma (FL). However, little information is available in the literature on the relationship between BCL2 protein, BCL2 gene status in FL and the patient outcomes. Also, understanding of IGH/BCL2 molecular rearrangement using real time PCR (RT-PCR), in follicular lymphoma in relation to survival might provide a more accurate and rational method of risk stratification to guide treatment and might suggest new therapeutic approaches as well. Methods: This study evaluated the relative frequency of t(14;18) by RT-PCR and its apoptosis-related BCL2 protein expression by an immunohistochemical assay (IHC) in fifty FL cases in tissues. In addition, we evaluated the relation of BCL2 protein expression to the translocation, together with the relation of both t(14;18) and BCL2 protein expression to the clinico-pathological features and survival data including progression free survival (PFS); and overall survival (OS) in order to evaluate their prognostic role in FL. Results & conclusion: There was a significant association of the t(14;18) with BCL2 protein expression, grading of FL, and the OS. In addition, there was a significant association of BCL2 protein expression with the grading of FL, OS, International Prognostic Index (IPI) score and performance status. However no significant association of t(14;18) or BCL2 protein expression with the other clinico-pathological features, and PFS. .
Up to 70% of thrombotic patients with no identifiable risk factors were termed idiopathic. Now, molecular diagnostics combined with existing laboratory techniques allow accurate classification of at least half of patients with inherited thrombotic disorders. However, the previously studied genetic variants explain only a fraction of all thromboembolic events, so the aim of the present study was to expand the genetic prevalence determination to include also more extensive thrombophilic gene polymorphisms in patients with DVT compared to healthy subjects in Egyptian population. This study was conducted on 75 patients with DVT and 45 age and sex matched healthy subjects as control group. The diagnosis of DVT was based on patient's history, clinical findings, D-dimer test, and confirmed by Doppler ultrasonography. Both groups were assessed for thrombophilic gene polymorphisms using multiplex polymerase chain reaction and reverse-hybridization technique through CVD strip assay. It was found that FV Leiden G1691A (P=0.001), Factor V H1299R (P=0.02), MTHFR A1298C (P=0.02), β-fibrinogen−455 G/A (P=0.01), PAI-1 4G/5G (P=0.03) and ACE (P=0.03) polymorphisms were all significantly associated with an increased risk of DVT. Our data are of extreme importance in clinical practice for introducing the extended CVD panel into routine molecular diagnostic tests to allow management of thrombotic patients and screening, thromboprophylaxis and genetic counselling for high-risk individuals..
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