CISD2 promotes lung squamous carcinoma cell migration and invasion via the TGF-B1-induced Smad2/3 signaling pathway

Background Although aberrant expression of CDGSH iron sulfur domain 2 (CISD2) contributes to the tumorigenesis and progression of numerous human cancers, the biological function of CISD2 and its specific prognostic value in lung squamous cell carcinoma (LUSC) have yet to be comprehensively explored. The current study aimed to elucidate the role of CISD2 in LUSC as well as the underlying molecular mechanisms. Methods Immunohistochemistry was conducted to detect the protein expression of CISD2 and analyze whether high expression of CISD2 affects the overall survival (OS) of LUSC patients. Cell proliferation, colony formation, wound healing and Transwell invasion assays were performed to clarify whether CISD2 contributes to LUSC cell proliferation and disease progression. Quantitative real-time reverse transcription-PCR and western blot assays were used to detect the levels of transcription factors and key epithelial-mesenchymal transition (EMT)-related markers in LUSC cells after CISD2 knockdown and overexpression to determine whether CISD2 regulates transforming growth factor-beta (TGF-β)-induced EMT in LUSC. Results Immunohistochemistry of human tissue microarrays containing 90 pairs of adjacent and cancerous tissues revealed that CISD2 is considerably overexpressed in LUSC and strongly linked to poor OS. Functional experiments suggested that silencing endogenous CISD2 inhibited the growth, colony formation, migration, and invasion of H2170 and H226 cell lines. Exogenous overexpression of CISD2 facilitated these phenotypes in SK-MES-1 and H2170 cells. Furthermore, CISD2 promoted EMT progression by increasing the expression of mesenchymal markers (N-cadherin, vimentin, Snail, and Slug) as well as SMAD2/3 and reducing the expression of the epithelial marker E-cadherin. Mechanistically, our studies provide the first evidence that CISD2 can promote EMT by enhancing TGF-β1-induced Smad2/3 expression in LUSC cells (AU)

The mechanism of VCP-mediated metastasis of osteosarcoma based on cell autophagy and the EMT pathway

Objective Study of the molecular mechanisms of metastasis is still the research focus for osteosarcoma (OS) prevention. This study investigates the mechanism of valosin-containing protein (VCP) promoting OS metastasis in vitro through autophagy and epithelial–mesenchymal transition (EMT). Methods Different cell lines of osteosarcoma (143B and MG63) were adopted in this study. The level of VCP expression in osteosarcoma cells was changed, and the level of autophagy and the progression of the epithelial–mesenchymal transition (EMT) were observed. Then autophagy and EMT in OS cells were changed artificially, and proliferation and migration ability were observed. Results The expression of LC3II/I was decreased, but the insolubilized P62 protein expression was increased in the VCP inhibiting group and the autophagy inhibitor treatment group. Simultaneously, E-cadherin protein expression increased while N-cadherin protein expression decreased in the VCP inhibiting group but increased in the TGF-β1 treatment group. In addition, suppressing VCP can cause a decrease in Transforming Growth Factor β1 (TGF-β1), smad2, smad3, phosphorylated smad2 (p-smad2), and phosphorylated smad3 (p-smad3). Autophagy inhibitors and agonists have no significant effect on the migration and invasion of OS cells but can significantly affect the ability of cells to resist anoikis. EMT inhibitors and agonists have a proportional effect on the migration and invasion of OS cells. Conclusion VCP is likely to promote the migration and invasion of OS cells by inducing EMT, possibly via TGF-β1/smad2/3 signaling pathway. In this process, VCP-mediated autophagy may contribute to successful distant metastasis of tumor cells indirectly (AU)

MiR-328-3p promotes TGF-B1-induced proliferation, migration, and inflammation of airway smooth muscle cells by regulating the PTEN/Akt pathway

Background: Recent studies have shown that the up-regulation of microRNA miR-328-3p expression increases seasonal allergy and asthma symptoms in children, but the specific mechanism remains unclear. Therefore, the aim of this study was to explore the role and mechanism of -miR-328-3p in transforming growth factor (TGF)-β1-induced airway smooth muscle cells (ASMCs). Methods: The effect of TGF-β1 on the expression of miR-328-3p in ASMCs was examined by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cells proliferation, migration, and inflammatory factors in TGF-β1-induced ASMCs were measured by cell counting kit-8 (CCK-8), transwell, and enzyme-linked immunosorbent assay (ELISA), respectively. Besides, TargetScan was used to predict phosphatase and tensin homolog (PTEN), the downstream target of miR-328-3p; double-luciferase reporter assay, western blot, and qRT-PCR were used to verify the targeting relationship between miR-328-3p and PTEN; western blot was also used to examine the effects of PTEN and miR-328-3p knockdown on the expression levels of PTEN, Akt, and p-Akt proteins. Results: The expression of miR-328-3p was up-regulated in TGF-β1-induced ASMCs. Knockdown of miR-328-3p significantly inhibited proliferation, migration, and inflammation of ASMCs induced by TGF-β1 and decreased levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β. The dual--luciferase reporter assay results confirmed that PTEN was a target gene of miR-328-3p. Moreover, inhibition of PTEN expression reversed the inhibitory effect of low miR-328-3p expression on -TGF-β1-induced ASMC’s proliferation, migration, and inflammation (AU)

CTRP3 regulates NF-kB and TGFB1/Smad3 pathways to alleviate airway inflammation and remodeling in asthmatic mice induced by OVA

Background: Asthma is a common illness with chronic airway inflammation. C1q/tumor necrosis factor (TNF)-related protein 3 (CTRP3) plays a vital role ininflammatory response, but its effect on asthma is imprecise. Herein, we analyzed the functions of CTRP3 in asthma. Methods: The BALB/c mice were randomized into four groups: control, ovalbumin (OVA), OVA+vector, and OVA+CTRP3. The asthmatic mice model was established by OVA stimulation. Overexpression of CTRP3 was implemented by the transfection of corresponding adeno-associated virus 6 (AAV6). The contents of CTRP3, E-cadherin, N-cadherin, smooth muscle alpha-actin (α-SMA), phosphorylated (p)-p65/p65, transforming growth factor-beta 1 (TGFβ1), and p-Smad3/Smad3 were determined by Western blot analysis. The quantity of total cells, eosinophils, neutrophils, and lymphocytes in bronchoalveolar lavage fluid (BALF) was assessed by using a hemocytometer. The contents of tumor necrosis factor-α and interleukin-1β in BALF were examined by enzyme-linked immunesorbent serologic assay. The lung function indicators and airway resistance (AWR) were measured. The bronchial and alveolar structures were evaluated by hematoxylin and eosin staining and sirius red staining. Results: The CTRP3 was downregulated in mice of OVA groups; however, AAV6-CTRP3 treatment markedly upregulated the expression of CTRP3. Upregulation of CTRP3 diminished asthmatic airway inflammation by decreasing the number of inflammatory cells and the contents of proinflammatory factors. CTRP3 markedly lessened AWR and improved lung function in OVA-stimulated mice. Histological analysis found that CTRP3 alleviated OVA-induced airway remodeling in mice. Moreover, CTRP3 modulated NF-κB and TGFβ1/Smad3 pathways in OVA-stimulated mice. Conclusion: CTRP3 alleviated airway inflammation and remodeling in OVA-induced asthmatic mice via regulating NF-κB and TGFβ1/Smad3 pathways (AU)

MicroRNA-34b-5p binds enhancer of zeste 2 to inhibit milk fat globule-EGF factor 8 expression, affecting liver fibrosis

Activated hepatic stellate cells (HSCs) are considered the major drivers in the process of hepatic fibrosis. This study intends to explore the mechanism underlying microRNA (miR)-34b-5p effects over liver fibrosis through the enhancer of zeste 2 (EZH2)/milk fat globule-EGF factor 8 (MFGE8) axis in HSCs. A liver fibrosis model was generated by carbon tetrachloride (CCl4) in C57BL/6 J mice and subjected to histological examinations and detection of HSC activation and miR-34b-5p/EZH2/MFGE8 expression. Primary HSCs were treated with transforming growth factor (TGF)-β and tested for proliferation, activation, and expression of fibrosis-related factors. A dual luciferase reporter assay was performed for confirming the targeted relationship between miR-34b-5p and EZH2. Chromatin immunoprecipitation was used to measure EZH2 enrichment in the MFGE8 promoter region. We found that miR-34b-5p was lowly expressed in the CCl4-induced mouse model. Overexpression of miR-34b-5p suppressed both TGF-β-induced HSC proliferation and the expression of fibrosis-related factors and HSC activation markers. A dual luciferase assay showed a binding relationship between miR-34b-5p and EZH2. Overexpression of miR-34b-5p reduced TGF-β-induced HSC activation by inhibiting EZH2 to promote MFGE8 expression. Overexpression of miR-34b-5p inhibited liver fibrosis in vivo through the EZH2/MFGE8 axis. Conclusively, overexpressing miR-34b-5p reduced TGF-β-induced HSC activation by inhibiting EZH2 and thereby promoting MFGE8 expression, and inhibited liver fibrosis in vivo through the EZH2/MFGE8 axis. (AU)

IL-10 and TGF-beta1 gene polymorphismsin Greek patients with recurrent aphthous stomatitis

Background: Recurrent aphthous stomatitis (RAS) is one of the most frequent inflammatory disorders of theoral mucosa. Cytokines, which play an important role in RAS pathogenesis, participate directly or indirectly innormal, immunological and inflammatory processes and are secreted from cells belonging to innate and adaptiveimmunity as a consequence of microbial and antigenic stimuli. Gene polymorphisms in specific cytokines maypredispose to RAS development. The aim of this study was the investigation and association of IL-10 and TGF-β1gene polymorphisms with RAS.Material and Methods: Study’s cohort consisted of 60 Greek patients diagnosed with RAS, including 40 patientswith minor, 10 patients with major and 10 with herpetiform aphthous ulcers. Forty age- and sex-matched controlsubjects were included in this study. DNA was extracted from whole blood samples of all patients and sequencespecific primers (SSP)-based polymerase chain reaction (PCR) was used for genotyping. Gene polymorphisms forcytokines IL-10 at loci -592 and -819 and for TGF-β1 at codon 10 were detected.Results: Significant differences between patients with minor RAS and healthy controls were recorded for IL-10genotypes distribution at position -592 (p=0.042) and -819 (p=0.045) with predominance of C/A and C/T genotypes in RAS patients, respectively. Also, in patients with minor and herpetiform aphthous ulcerations, heterozygousTGF-β1 genotype C/T at codon 10 was associated with increased risk of RAS (p=0.044 and p=0.020, respectively).Conclusions: These data provide evidence that genetic predisposition for RAS and possibly its specific clinical variants is related with the presence of gene polymorphisms for specific cytokines, including IL-10 and TGF-β1, which,in turn, may vary according to geographic origin and genetic background. (AU)

Correlation analysis of TGF-β1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α in refractory chronic rhinosinusitis: A retrospective study

Objective: To investigate the potential correlation of transforming growth factor-β (TGF-β), matrix metalloprotein 9 (MMP-9), tissue inhibitor of metalloproteinases 1 (TIMP-1), Interleukin 1 (IL-1), IL-4, IL-6, IL-17, and tumor necrosis factor alpha (TNF-α) in refractory chronic rhinosinusitis.Methods: A total of 150 participants were retrospectively included in this study from August 2018 to February 2020. The people enrolled were equally allocated into refractory group (patients with refractory chronic rhinosinusitis), chronic group (patients with chronic rhinosi-nusitis), and control group (normal people). The level of TGF-β1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α were recorded. The unconditional multivariate binary logistic regression was used to analyze the factors affecting refractory chronic rhinosinusitis.Results: The Davos score, T&T olfactometer threshold test, and Lund-Mackay CT scores in refractory group were significantly higher than the chronic group (P<0.05). The level of TGF-β1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α in the refractory group were significantly higher than the chronic group and the control group (all P<0.05). Similarly, the level of the above mentioned indexes in the chronic group were significantly higher than the control group (P<0.05). The Davos score, T&T olfactometer threshold test score, Lund-Mackay CT score, and the level of TGF-β1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α positively correlated with refractory chronic rhinosinusitis. Moreover, the unconditional multivariate binary logistic regression showed that the influencing factors of refractory chronic rhinosinusitis included TGF-β1, MMP-9, TIMP-1, IL-1, IL-4, IL-6, IL-17, and TNF-α.Conclusion: The findings of the present study provide evidence for TGF-β1, MMP-9, TIMP-1, IL-4, IL-6, IL-17, and TNF-α as the influencing factors of refractory chronic rhinosinusitis (AU)

Dexmedetomidine represses TGF-beta1-induced extracellular matrix production and proliferation of airway smooth muscle cells by inhibiting MAPK signaling pathway

Background Airway remodeling is implicated in the pathogenesis of asthma, and abnormal proliferation of airway smooth muscle cells (ASMCs) contribute to airway remodeling. Inflammatory mediator, transforming growth factor-β1 (TGF-β1), stimulates the proliferation of ASMCs, and is associated with airway remodeling in asthma. Dexmedetomidine (DEX) has been widely used in the adjuvant therapy of acute asthma.Objective The potential effects of DEX on extracellular matrix (ECM) production and proliferation of ASMCs were investigated in this study.Material and Methods Human ASMCs were incubated with TGF-β1 for 48 hours, and then treated with different concentrations of DEX for another 24 hours. Cell proliferation was detected by MTT and BrdU (5’-bromo-2’-deoxyuridine) staining. Flow cytometry was used to assess cell apoptosis, and western blot was applied to identify the underlying mechanism.Results TGF-β1 induced increase in cell viability and bromodeoxyuridine (BrdU) positive cells in ASMCs while repressed cell apoptosis. Second, TGF-β1-induced ASMCs were then treated with different concentrations of DEX. Cell viability of TGF-β1-induced ASMCs was decreased by incubation of DEX. The number of BrdU positive cells in TGF-β1-induced ASMCs was reduced by incubation of DEX. Moreover, incubation of DEX promoted cell apoptosis of TGF-β1-induced ASMCs. Third, incubation of DEX attenuated TGF-β1-induced increase in fibronectin, collagen I, MMP9, and versican in ASMCs. Lastly, the up-regulation of phosphorylated extracellular receptor kinase (p-ERK), phosphorylated Jun N-terminal Kinase (p-JNK), and p-p38 in TGF-β1-induced ASMCs was reversed by incubation of DEX.Conclusion DEX suppressed TGF-β1-induced ECM production and proliferation of ASMCs through inactivation of p38 mitogen-activated protein kinase (MAPK) pathway, providing a potential strategy for prevention of asthma (AU)

6-Gingerol, a functional polyphenol of ginger, reduces pulmonary fibrosis by activating Sirtuin1

Pulmonary fibrosis in general is the final common outcome of various interstitial lung diseases. In recent years, the incidence of pulmonary fibrosis has been rising with poor prognosis. 6-gingerol is deemed as a functional polyphenol of ginger. The aim of the present study was to investigate the effect of 6-gingerol, on pulmonary fibrosis. Mice were randomly divided into four groups: control, bleomycin, bleomycin + 6-gingerol 100 mg/kg, bleomycin + 6-gingerol 250 mg/kg, and the survival rates of the groups were recorded. Pathological and fibrotic changes in the lungs were identified by H&E and Masson staining, respectively. The levels of hydroxyproline and protein deposited in lung tissues were then, respectively, determined by colorimetry and western blotting. Subsequently, the proportion of cells and inflammatory factors in the alveolar lavage fluid were estimated. Following the identification of the possibility of Sirtuin1 (SIRT1) in the pharmacological mechanism through molecular docking and western blotting, human embryonic lung fibroblasts MRC-5 were treated with TGF-β1 and SIRT1 inhibitor to study the role of SIRT1 in the regulatory effect of 6-gingerol. From the results, 6-gingerol was found to increase the survival rate of mice and reduce lung pathology and fibrosis in mice. And, it significantly reduced the levels of hydroxyproline and the proteins deposited in lung tissues. Moreover, the number of neutrophils, basophils, monocytes, and the levels of inflammatory factors in the alveolar lavage fluid were also reduced. SIRT1 inhibitor blocked the function of 6-gingerol to inhibit fibrosis. To sum up, 6-gingerol relieves pulmonary fibrosis via activating SIRT1. This finding expands the pharmacological effect of 6-gingerol, and it is expected to advance the development of treatments for pulmonary fibrosis (AU)

Bone pain management with opioid medication in a patient with Camurati-Engelmann disease: a case report / Manejo del dolor óseo con medicación opioide en una paciente con enfermedad de Camurati-Engelmann: reporte de un caso

Introduction: Camurati-Engelman Disease is a rare genetic sclerosing bone dysplasia with periosteal and endosteal thickening of the cortical of the long bones. It generates pain secondary to the reduction of the medullary canal that is usually controlled with corticosteroids and, in severe cases, with surgical decompression. Case history: We present the case of a woman with a genetic diagnosis of Camurati-Engelman Disease with poor pain control with corticosteroid management and surgical procedures throughout her childhood and early adulthood. In whom optimal pain control was achieved with pain regimen with hydrocodone analgesic management. This is the first case described in the literature for adequate pain control using an opioid drug. Discussion: CE disease is an extremely rare genetic entity with little more than 300 cases reported in the world. It is generated by an alteration in the gene for growth factor-beta 1 (TGF-B1); it has a varied clinical presentation that can begin with bone alterations accompanied by muscle weakness, joint angular alterations, headache, and nerve compressions. It has a differential diagnosis with some genetic entities that may present clinical similarity, but its morphological and radiological characteristics are distinctive. The usual management of bone pain generated by this entity is based on corticosteroids, in addition to losartan or surgical intervention aimed at reducing cortical changes. The intervention with opioid analgesics accompanied by a rehabilitation plan is not a frequent report, this being a case of success due to the refractoriness of the symptoms in a patient with chronic pain, with a positive impact on her functionality and quality of life. Conclusion: It is considered that analgesic management with opioids may be a treatment option in patients with Camurati-Engelman disease refractory to corticosteroid management and surgical interventions.(AU)

Introducción: La enfermedad de Camurati-Engelman (CE) es una displasia ósea esclerosante rara, de causa genética. Se presenta con engrosamiento perióstico y endóstico de la cortical de los huesos largos. Genera dolor secundario a la reducción del canal medular, que habitualmente se controla con corticoides y en casos severos, con descompresión quirúrgica. Historia del caso: Presentamos el caso de una mujer con diagnóstico genético de enfermedad de Camurati-Engelman, con mal control del dolor, con manejo de corticosteroides y procedimientos quirúrgicos a lo largo de su niñez y adultez temprana. Se logró un control óptimo del dolor con un régimen con manejo analgésico con hidrocodona. Este es el primer caso descrito en la literatura de un adecuado control del dolor con un medicamento opioide. Discusión: La enfermedad de CE es una entidad genética extremadamente rara, con poco más de 300 casos reportados en el mundo. Se genera por una alteración en el gen del factor de crecimiento beta 1 (TGF-B1). Tiene una presentación clínica variada que puede iniciar con las alteraciones óseas acompañado de debilidad muscular, alteraciones angulares articulares, cefalea y compresiones nerviosas. Tiene diagnóstico diferencial con algunas entidades genéticas que pueden presentar similitud clínica, pero su característica morfológica y radiológica es distintiva. El manejo usual del dolor óseo generado por esta entidad se basa en corticoesteroides, además de losartán o intervenciones quirúrgicas orientadas a disminuir los cambios corticales. La intervención con analgésicos opioides, acompañada de un plan de rehabilitación, no es un reporte frecuente, siendo este un caso de éxito ante la refractariedad de los síntomas en una paciente con dolor crónico, impactando de manera positiva en su funcionalidad y calidad de vida.(AU)

Neutral endopeptidase depletion decreases colon cancer cell proliferation and TGF-B1 synthesis in indirect co-cultures with normal colon fibroblasts

Purpose Although recent studies have suggested that neutral endopeptidase (NEP) is implicated in the regulation of colon cancer (CC) cell growth and metastasis, the influence of the tumor microenvironment on this role of NEP has not been investigated so far. Normal colon fibroblasts (NCFs) constitute a component of the stroma surrounding a tumor in an early stage of its development. NCFs can influence transformed cells via different paracrine factors, including TGF-β1. This in vitro study was undertaken to evaluate the role of NEP in CC promotion in conditions of indirect co-culture of CC cells (LS180 and SW620) with NCFs (CCD-18Co) or their conditioned medium (CM-18Co). Methods We examined cell proliferation (with the BrdU assay) and invasiveness (using BME-coated inserts, 8 µm) of NEP-expressing, NEP-silenced (siRNA), and NEP-inhibited (with thiorphan, i.e. a NEP specific inhibitor) CC cells cultured alone or co-cultured with CCD-18Co or with their conditioned medium. The Western blot and ELISA methods were used to assess the level of TGF-β1. Results The results showed that the co-culture of the NEP-depleted CC cells with NCFs or their conditioned medium resulted in a significant decrease in cell proliferation in comparison with the proliferative potential of NEP-silenced/inhibited CC cells cultivated alone. In contrast, the NEP depletion did not influence the invasiveness of CC cells in the co-cultures. The co-culture of CC cells with CCD-18Co or CM-C18Co resulted in increased synthesis of TGF-β1, while the NEP downregulation decreased the synthesis of TGF-β1 in CC cells and abolished the stimulatory effect of the co-cultures on TGF-β1 production. Conclusions The results suggest that the expression of NEP by colon cancer cells is essential for their proliferation and TGF-β1 synthesis during paracrine interactions with NCFs (AU)

Capacitación osteogénica in vitro de células madre mesenquimales de médula ósea para su aplicación en resecciones segmentarias de hueso / Osteogenic in vitro training of bone marrow mesenquimal cells for application in segmentary bone resections

OBJETIVO: Obtener continuidad ósea en un modelo experimental de resección segmentaria en la diáfisis del fémur mediante tratamiento con células mesenquimáticas indiferenciadas comprometidas al linaje osteogénico. MATERIAL Y MÉTODO: Se obtuvieron células mesenquimáticas indiferenciadas a partir de médula ósea de ratas Wistar singénicas, se diferenciaron al linaje osteogénico y se embebieron en bloques de hidroxiapatita. Se implantaron en una resección segmentaria en la diáfisis del fémur, que se sintetizó con una placa de 1,5mm de grosor. Se calcularon distribuciones binomiales estableciéndose un grupo experimental y 3 de control, constituidos por 8 elementos cada grupo. Grupo I, relleno con aloinjerto; grupo II, con hidroxiapatita; grupo III, con hidroxiapatita embebida con células osteocomprometidas; grupo IV con células osteocomprometidas mediante cultivo tridimensional. Se realizó estadística descriptiva con distribución de frecuencias mediante la prueba de Fisher, considerándose significativo el valor de p < 0,05. RESULTADOS: El grupo I presentó buena consolidación, sin rotura de placas. El grupo II mostró tejido fibroso y rotura de todas las placas. El grupo III mostró tejido óseo, pero en todos los casos se rompieron las placas. El grupo IV mostró consolidación sin rotura de placas. CONCLUSIÓN: La terapia mediante células mesenquimáticas indiferenciadas en cultivos tridimensionales produce tejido óseo y asegura una estabilización mecánica permanente. Limitaciones: antes de la inferencia humana es necesario realizar el experimento en grupos con más elementos

OBJECTIVE: To achieve bone continuity in an experimental model of segmental resection of femur bone by applying a treatment with committed to osteogenic bone linage mesenchymal stem cells. MATERIAL AND METHOD: Bone marrow mesenchymal stem cells, obtained from syngeneic Wistar murine, were committed into osteogenic lineage and embedded within a hydroxipatite block. They were implanted in an experimentally created diaphyseal femur resection model. The diaphysis was synthetized with a 1.5mm thick plate. In order to calculate binomial distributions, we stablished one experimental and 3 control groups of 8 elements each: Group I, filling the gap with allograft; group II, filling with a hydroxyapatite block without cells; group III, filling with the hydroxyapatite block embedded with committed cells, and group IV, with the hydroxyapatite embedded with osteoinduced cells in a 3 dimensions TRAP culture. Descriptive analysis was performed by frequency distribution and Fisher statistic test. Level of statistical significance was considered at P<.05. RESULTS: Group I presented good bone consolidation and no plate breakage. Group II showed fibrous but non-bone tissue, with rupture of all plates. Group III showed bone tissue in all cases, but the plates broke in all of them, while in group IV bone consolidation was achieve without any plate rupture. CONCLUSION: Cell therapy with mesenchymal stem cells, trained in a 3 dimensions cell culture, produces bone tissue and ensures the permanence of the mechanical stabilization performed in a segmental resection model. Limitations: A study with a larger sample size is necessary before planning the human inference

Atopy can be an interfering factor in genetic association studies of beta-lactam allergy

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Budesonide up-regulates vitamin D receptor expression in human bronchial fibroblasts and enhances the inhibitory effect of calcitriol on airway remodeling

Introduction and objectives: Transforming growth factor β1 (TGFβ1) and dysregulated microRNA-21 (miR-21) expression is associated with TGFβ/Smad signaling pathway activation and fibrosis. While calcitriol has been shown to improve airway remodeling in asthmatic mice, its mechanism remains unknown. In this study, the effect of calcitriol on the TGFβ/Smad signaling pathway and miR-21 expression in human bronchial fibroblasts was investigated to explore the mechanism of action of calcitriol and the inhaled glucocorticoid, budesonide, in airway remodeling. Materials and methods: Human bronchial fibroblasts were pretreated with budesonide, calcitriol, or budesonide plus calcitriol, and stimulated with TGFβ1 for 48h. Quantitative real-time PCR was used to determine the expression of miR-21. Western blot was used to determine airway remodeling-related proteins, TGFβ/Smad signaling pathway-related proteins, glucocorticoid receptor, and vitamin D receptor (VDR) expression. Results: Both budesonide and calcitriol down-regulated miR-21 expression in human bronchial fibroblasts, up-regulated Smad7 expression, and inhibited the expression of airway remodeling-related proteins. Both budesonide and calcitriol up-regulated the low expression of VDR induced by TGFβ1 in human bronchial fibroblasts. The expression of VDR in the combined treatment group (budesonide plus calcitriol) was significantly higher than that in the calcitriol treatment group. The expression of collagen type I in the combined treatment group was significantly lower than that in the calcitriol treatment group. Conclusions: Calcitriol can up-regulate the expression of VDR in human bronchial fibroblasts and exert an anti-airway remodeling effect. Budesonide can up-regulate the expression of VDR in human bronchial fibroblasts and enhance the inhibitory effect of calcitriol on airway remodeling

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Mecanismos patogénicos en la fibrosis pulmonar idiopática. Implicación de los canales de aquoporina 1 (aqp1) / Pathogenic mechanisms in idiopathic pulmonary fibrosis. Involvement of aquoporin 1 (AQP1) channels

La fibrosis pulmonar idiopática (FPI) es una enfermedad pulmonar intersticial difusa (EPID) y progresiva. La FPI resulta de la alteración en la reepitelización tras la lesión de las células epiteliales alveolares. Se produce un aumento en la apoptosis epitelial y en la síntesis de mediadores profibróticos, con la consiguiente proliferación de fibroblastos, transformación a miofibrobastos y el depósito incontrolado de matriz extracelular. La aquoporina 1 (AQP1), es una proteína que facilita el movimiento de agua entre el espacio aéreo pulmonar y el parénquima y se ha demostrado que se regula al alza en animales expuestos a hipoxia. La AQP1 se expresa en células endoteliales, pero no en el epitelio alveolar pulmonar. Más recientemente se ha asociado a la AQP1 en los mecanismos implicados en la proliferación celular. Nuestro objetivo principal ha sido evaluar la participación de las Aquoporinas (AQPs) pulmonares en la patogenia de la FPI. Hemos analizado la expresión de AQP1 en biopsias de pacientes diagnosticados con FPI según los criterios de la ATS/ERS/ALAT de 2011 y en otras patologías pulmonares tales como la neumonitis por hipersensibilidad, sarcoidosis y biopsias de controles sanos. Los resultado de la inmunohistoquímica revelaron una intensa expresión de AQP1 en neumocitos tipo II hiperplásicos solo en las muestras obtenidas de pacientes con FPI. Además hay una inducción de la expresión de AQP1 (mRNA y proteína) tras la estimulación con TGF-ß lo que acompaña a los cambios típicos del proceso de transición epitelio mesenquimal. Por lo tanto, la aparición de AQP1 en las células hiperplásicas tipo II y su regulación podrían estar implicadas en la patogénesis de la FPI

Idiopathic pulmonary fibrosis (IPF) is a diffuse interstitial lung disease (ILD) that is progressive. IPF results from altered re-epithelialization after injury to alveolar epithelial cells. There is an increase in epithelial apoptosis and synthesis of pro-fibrotic mediators, with consequent proliferation of fibroblasts, transformation into myofibroblasts and uncontrolled deposition of extracellular matrix. Aquoporin 1 (AQP1) is a protein that facilitates the movement of water between the pulmonary airspace and the parenchyma and has been shown to be upregulated in animals exposed to hypoxia. AQP1 is expressed in endothelial cells, but not in the pulmonary alveolar epithelium. More recently, AQP1 has been associated in the mechanisms involved in cell proliferation. Our primary objective has been to assess the participation of lung aquoporins (AQPs) in the pathogenesis of IPF. We have analyzed the expression of AQP1 in biopsies of patients diagnosed with IPF according to the ATS/ERS/ALAT 2011 criteria and in other lung diseases such as hypersensitivity pneumonitis, sarcoidosis and biopsies of healthy controls. Immunohistochemistry results revealed an intense expression of AQP1 in Type II pneumocyte hyperplasia only in samples obtained from patients with IPF. Additionally, AQP1 (mRNA and protein) expression is induced after stimulation with TGFß, which accompanies typical changes in the mesenchymalepithelial transition process. Therefore, the appearance of AQP1 in Type II cell hyperplasia and its regulation could be involved in the pathogenesis of IPF

Correlación de niveles del factor de crecimiento transformante Beta-1 con severidad de vitreorretinopatía proliferativa en pacientes con desprendimiento de retina regmatógeno / Correlation of transforming growth factor Beta-1 vitreous levels with clinical severity of proliferative vitreoretinopathy in patients with rhegmatogenous retinal detachment

Objetivo: Correlacionar la concentración vítrea del factor de crecimiento transformante Beta-1 (TGFBeta-1), con el grado de severidad clínica de la vitreorretinopatía proliferativa (VRP). Diseño: Se realizó un estudio prospectivo, observacional, transversal de casos y controles. Participantes: Se incluyeron 40 pacientes con diagnóstico de VRP secundaria a desprendimiento de retina regmatógeno. Métodos: Se obtuvo vítreo en pacientes sometidos a vitrectomía pars plana por desprendimiento de retina regmatógeno, que fueron atendidos durante el periodo de agosto del 2015 a junio del 2016, en un centro de referencia nacional para la atención oftalmológica en la Ciudad de México, México. Se cuantificaron los niveles de TGFBeta-1 mediante técnica de ELISA. Se realizó una prueba ANOVA para la comparación de los distintos grupos junto con una prueba de Dunns post hoc. Se consideró una diferencia estadísticamente significativa al obtener p < 0,05. Resultados: Se cuantificaron los niveles de TGFBeta-1 y se encontraron las siguientes medias para cada grupo. En el grupo con VRP grado A, 1150,6 ± 452,08 pg/ml, VRP grado B: 1129,6 ± 365,54 pg/ml y VRP grado C: 1146,4 ± 330,21 pg/ml. El análisis estadístico no encontró diferencias significativas cuando se comparan los diferentes grupos de VRP (p = 0,53). Sin embargo, al realizar el análisis diferencial para cada grado de severidad, se observó un aumento estadísticamente significativo en la expresión de TGFBeta-1 en el grupo de pacientes con VRP-A, en relación con mayor número de días de evolución del desprendimiento (p = 0,03). Diferencias que no fueron estadísticamente significativas para VRP-B y C (p = 0,16) (p = 0,16). Conclusión: El comportamiento del TGFβ-1 no guardó relación directa con la severidad clínica, esto sugiere que debe haber otros factores involucrados en las etapas avanzadas de VRP, mientras que podría ser que el TGFβ-1 tenga mayor relevancia durante las etapas iniciales de la evolución clínica promoviendo la transición epitelial-mesenquimatosa debido a su mayor expresión en las etapas iniciales. De lo cual se puede concluir que cada isoforma desempeña un papel muy particular en el complejo proceso de la VRP

Objective: To correlate the vitreous concentration of transforming growth factor β-1 (TGF Beta-1) with the degree of clinical severity of proliferative vitreoretinopathy (PVR). Design: A prospective, observational, cross-sectional study carried out on cases and controls. Participants: The study included 40 patients with a diagnosis of PVR secondary to rhegmatogenous retinal detachment. Methods: Vitreous was obtained in patients undergoing pars plana vitrectomy by rhegmatogenous retinal detachment, who were treated during the period from August 2015 to June 2016, in a national reference centre for ophthalmological care in Mexico City, Mexico. The levels of TGFBeta-1 were quantified by ELISA technique. An ANOVA test was performed for the comparison of the different groups, together with a post-hoc Dunns test. A statistically significant difference was considered when obtaining P <.05. Results: The levels of TGFBeta-1 were quantified, and the following means were found for each group: In the group with PVR grade A, 1150.6 ± 452.08 pg / ml, PVR grade B: 1129.6 ± 365.54 pg / ml, and PVR grade C: 1146.4 ± 330.21 pg / ml. The statistical analysis did not find significant differences when comparing the different PVR groups. (P=.53). However, when performing the differential analysis for each level of severity, a statistically significant increase in the expression of TGFBeta-1 was observed in the group of patients with PVR-A at a greater number of days of evolution of the detachment. (P=.03). There were no statistically significant differences for PVR-B and PVR-C (P=.16 and P=.16, respectively). Conclusion: Although the levels of TGFBeta-1 are not directly related to the clinical severity grade, suggesting that there must be other factors involved in the advanced stages of PVR, TGFBeta-1 may have greater relevance during the initial stages of the clinical course by promoting the epithelial-mesenchymal transition due to its greater expression in PVR-A. Thus, it can be concluded that each isoform plays a very particular role in the complex process of PVR

Single nucleotide polymorphisms of the genes encoding IL-10 and TGF-Beta1 in Iranian children with atopic dermatitis

Background: Atopic dermatitis is an inflammatory skin disease in which both genetic and environmental factors interact to determine the susceptibility and severity of the disease. Objective: The aim of this study was to determine the association between atopic dermatitis and IL-10 and TGF-Beta1 gene polymorphisms. Methods: The allele and genotype frequencies of genes encoding for IL-10 and TGF-Beta1 were investigated in 89 patients with atopic dermatitis in comparison with 138 in the control group using the PCR-SSP method. Results: A significant increase was found in the frequency of the TGF-Beta1 codon 10/C allele among patients (p < 0.001, OR = 6.77), whereas a significant decrease was observed in the frequency of the T allele at the same position (p < 0.001, OR = 0.14). The frequency of the TGF-Beta1 codon 25/G allele in the control group was significantly higher than among patients (p < 0.001, OR = 0.08). A significant positive correlation was seen between CC (p < 0.001, OR = 15.10) and CG (p < 0.001) genotypes and AD at codons 10 and 25, respectively. The most frequent haplotypes among patients was TGF-Beta1 CG which was significantly higher than in the control subjects (50% in patients vs. 39.9% in controls, p = 0.042). A significant increase was found in the frequency of TGF-Beta CC (36% in patients vs. 7.6% in controls, p < 0.001) and TC (14% in patients vs. 0% in controls, p < 0.001) haplotypes among patients compared to controls. By contrast, the TGF-Beta1 TG haplotype was significantly lower in patients than controls (0% in patients vs. 52.5% in controls, p < 0.001). There were no significant differences in the frequency of alleles, genotypes and haplotypes of the IL-10 gene. Conclusions: We found a strong association between the polymorphisms of the TGF-Beta1 gene at codon 10 and codon 25 positions and atopic dermatitis (AU)

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Valor de los niveles urinarios de interleucina 6, factor de crecimiento epidérmico, proteína quimioatractante de monocitos de tipo 1 y factor de crecimiento transformante Beta1 para la predicción de la extensión de las lesiones de fibrosis en biopsias de enfermos con nefropatía IgA / Value of urinary levels of interleukin-6, epidermal growth factor, monocyte chemoattractant protein type1 and transforming growth factor Beta1 in predicting the extent of fibrosis lesions in kidney biopsies of patients with IgA nephropathy

Objetivo: Analizar las asociaciones entre el nivel urinario de IL-6, EGF, MCP-1 y TGFβ1 y las características clínicas, bioquímicas y anatomopatológicas en enfermos con nefropatía IgA primaria y determinar su capacidad para realizar una estimación de la extensión de las lesiones de esclerosis glomerular e intersticial. Pacientes y métodos: Se estudió a 58 enfermos con nefropatía IgA. Se determinaron los niveles urinarios de IL-6, EGF, MCP-1 y TGFβ1 en el momento del diagnóstico. Tras realizar un análisis de la extensión de las lesiones renales mediante morfometría cuantitativa y mediante los criterios de Oxford, se analizó la capacidad de dichas moléculas para estimar la extensión de las lesiones glomerulares e intersticiales de fibrosis. Resultados: La IL-6, MCP-1 y TGF-β1 se asociaron a glomeruloesclerosis focal y a la extensión de la fibrosis intersticial, pero no a la presencia de proliferación mesangial, intracapilar o extracapilar. EGF presentó una asociación negativa con la fibrosis intersticial. Al categorizar a los enfermos según la clasificación de Oxford, los enfermos con scores T1 y T2 presentaron niveles significativamente superiores de IL-6, MCP-1 y TGFβ1, y niveles de EGF significativamente inferiores que los enfermos con T0. Tanto mediante regresión múltiple como mediante regresión logística, los niveles de MCP-1, IL-6 y EGF fueron predictores independientes de la superficie de fibrosis, tras ajustar por edad y FGe. Conclusión: La determinación de la concentración urinaria de IL-6, EGF y MCP-1 proporciona una información adicional que mejora de forma significativa la estimación de la superficie de fibrosis intersticial (AU)

Objective: To analyse the associations between urinary levels of IL-6 EGF, MCP-1 and TGFβ1 and clinical, biochemical and histopathological characteristics in patients with primary IgA nephropathy and their ability to predict the extent of lesions of glomerular and/or interstitial sclerosis. Patients and methods: A total of 58 patients with IgA nephropathy were studied. We determined the urine levels of IL-6, EGF, MCP-1, and TGFβ1 at the time of diagnosis. The extent of glomerular and interstitial fibrosis was analyzed by quantitative morphometry and kidney biopsies were classified according to the Oxford criteria. We analysed the ability of these molecules to predict the extent of glomerular and interstitial fibrosis lesions. Results: IL-6, TGFβ1 and MCP-1 were associated with focal glomerulosclerosis and interstitial fibrosis extension but not with the presence of mesangial, extracapillary or endocapillary proliferation. EGF showed a negative association with interstitial fibrosis. By categorising patients according to the Oxford classification, patients with T1 and T2 scores had significantly higher levels of IL-6, MCP-1, TGF-β1 and significantly lower levels of EGF than patients with T0 scores. By multiple regression and logistic regression analyses, the levels of MCP-1, IL-6 and EGF were independent predictors of the fibrosis surface, after adjusting for age and eGFR. Conclusion: The urinary concentration of IL-6, EGF and MCP-1 provides additional information that significantly improves the estimation of the surface of interstitial fibrosis in patients with IgA nephropathy (AU)

Polymorphisms of TGFB1, TLE4 and MUC22 are associated with childhood asthma in Chinese population

Objective: To investigate whether the genetic variants of TGFB1, TLE4, MUC22 and IKZF3 are associated with the development of asthma in Chinese children. Methods: 572 adolescent asthma patients and 590 age-matched healthy controls were included in this study. A total of four SNPs were genotyped, including rs2241715 of TGFB1, rs2378383 of TLE4, rs2523924 of MUC22, and rs907092 of IKZF3. Allele frequencies of the patients and the control group were compared by the Chi-square test. The Student t test was used to analyse the relationship between genotypes and clinical feature of the patients. Results: Patients were found to have significantly different frequencies of allele A of rs2241715, allele G of rs2378383 and allele A of rs2523924 as compared with the controls (40.4% vs. 45.9%, p = 0.01 for rs2241715; 17.2% vs. 13.4%, p=0.01 for rs2378383; 15.3% vs. 11.9%, p = 0.02 for rs2523924). For patients with severe asthma, those with genotype AA/AG of rs2241715 had remarkably higher FEV1% as compared with those with genotype GG (59.1 ± 4.3% vs. 55.4 ± 3.7%, p < 0.001). Moreover, those with genotype GG/GA of rs2378383 had remarkably lower FEV1% as compared with those with genotype AA (54.6 seg ± 2.9% vs. 58.6 ± 4.1%, p < 0.001). Conclusions: Genes TGFB1, TLE4 and MUC22 are associated with the risk of childhood asthma in Chinese population. Our results associating TGFB1 and TLE4 with clinical features of asthma suggest potential application of these parameters in the management of asthma children (AU)

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Comparative immunoexpression of ICAM-1, TGF-Beta1 and ki-67 in periapical and residual cysts

BACKGROUND: This study compared the immunohistochemical expression of ki-67, transforming growth factor beta 1 (TGF-Beta 1) and intercellular adhesion molecule-1 (ICAM-1) in inflammatory periapical cysts and residual cysts. MATERIAL AND METHODS: The study sample was composed by 25 periapical cysts and 25 residual cysts and immunohistochemical reactions were carried out using antibodies directed against ICAM-1, TGF-β1 and ki- 67. Clinical, radiological, gross, histological and immunohistochemical data were tabulated for descriptive and comparative analysis using the SPSS software and differences were considered statistically significant when p < 0.05%. RESULTS: There were no differences between the expression of ICAM-1 (p = 0.239) and TGF-β1 (p = 0.258) when comparing both groups. Ki-67 labeling index was higher in residual cysts compared to periapical cysts (p = 0.017). CONCLUSIONS: Results from the present study suggest that some specific inflammatory stimuli on residual cysts would modulate their mechanisms of etiopathogenesis, growing and repair