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Long non-coding RNAs (lncRNAs) play a significant biological role in the regulation of various cellular processes such as cell proliferation, differentiation, apoptosis and migration. In various malignancies, lncRNAs interplay with some main cancer-associated signaling pathways, including the Hippo signaling pathway to regulate the various cellular processes. It has been revealed that the cross-talking between lncRNAs and Hippo signaling pathway involves in gastrointestinal (GI) cancers development and progression. Considering the clinical significance of these lncRNAs, they have also been introduced as potential biomarkers in diagnostic, prognostic and therapeutic strategies in GI cancers. Herein, we review the mechanisms of lncRNA-mediated regulation of Hippo signaling pathway and focus on the corresponding molecular mechanisms and clinical significance of these non-coding RNAs in GI cancers.
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Burns injuries are prone to hospital-acquired infections, and Pseudomonas aeruginosa is one of the most common causes of mortality and morbidity in patients with burn injuries. Thus, this study aimed to analyze the effects of topical treatment with bone marrow (BM-MSC) and adipose mesenchymal stem cells (AD-MSC) encapsulated in collagen and fibrin scaffolds in a Balb/c model of burn wound infection. Extraction of stem cells from adipose and bone marrow tissue of rats was performed and cells were characterized using standard methods. Then, collagen, fibrin and collagen-fibrin scaffolds were constructed and the extracted cells were encapsulated in all three scaffolds. Then, 3rd degree burn was induced in mice and 1.5 × 108 (CFU/ml) of P. aeruginosa was introduced to the burn wound. Subsequently, after 24 h of inducing wound infection, encapsulated MSCs were introduced as dressings to burn wound infection and microbial load as well as rate of wound infection healing was measured. The results of this study showed that the use of BM-MSC and AD-MSC encapsulated in collagen-fibrin scaffold reduced the bacteria load down to 54 and 21 CFU/gr, respectively (P < 0.05). Moreover, BM-MSC and AD-MSC encapsulated in collagen-fibrin showed 80% and 75% wound healing, respectively (P < 0.05). Also, we found no significant between cell origin and healing. Encapsulation of MSCs into collagen-fibrin scaffolds could be effective not only against P. aeruginosa infection, but also healing and regeneration of burn wound.
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Queimaduras , Células-Tronco Mesenquimais , Infecção dos Ferimentos , Humanos , Ratos , Camundongos , Animais , Pseudomonas aeruginosa , Hidrogéis/uso terapêutico , Medula Óssea , Fibrina/uso terapêutico , Queimaduras/tratamento farmacológico , Cicatrização , Colágeno/uso terapêutico , Antibacterianos/uso terapêutico , Infecção dos Ferimentos/terapia , Administração Tópica , Células da Medula ÓsseaRESUMO
Introduction: Human endometrial mesenchymal stem cells (hEnMSCs) are a rich source of mesenchymal stem cells (MSCs) with multi-lineage differentiation potential, making them an intriguing tool in regenerative medicine, particularly for the treatment of reproductive and infertility issues. The specific process of germline cell-derived stem cell differentiation remains unknown, the aim is to study novel ways to achieve an effective differentiation method that produces adequate and functioning human gamete cells. Methods: We adjusted the optimum retinoic acid (RA) concentration for enhancement of germ cell-derived hEnSCs generation in 2D cell culture after 7 days in this study. Subsequently, we developed a suitable oocyte-like cell induction media including RA and bone morphogenetic protein 4 (BMP4), and studied their effects on oocyte-like cell differentiation in 2D and 3D cell culture media utilizing cells encapsulated in alginate hydrogel. Results: Our results from microscopy analysis, real-time PCR, and immunofluorescence tests revealed that 10 µM RA concentration was the optimal dose for inducing germ-like cells after 7 days. We examined the alginate hydrogel structural characteristics and integrity by rheology analysis and SEM microscope. We also demonstrated encapsulated cell viability and adhesion in the manufactured hydrogel. We propose that in 3D cell cultures in alginate hydrogel, an induction medium containing 10 µM RA and 50 ng/mL BMP4 can enhance hEnSC differentiation into oocyte-like cells. Conclusion: The production of oocyte-like cells using 3D alginate hydrogel may be viable in vitro approach for replacing gonad tissues and cells.
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Introduction: The relation between physical well-being and chronic pain is complex and involves several subjective and objective covariates. We aimed to assess the role of mediator, confounder, or interactor played by covariates, including sleep quality, physical activity, perceived stress, smoking, and alcohol drinking in the relation between physical well-being and chronic pain. Method: We used Poisson regression to obtain incidence rate ratios (IRR) of the association between physical well-being and chronic pain in a cohort study carried out among university students. We applied General Structural Equation Modeling (GSEM) to assess mediation and stratum-specific analyses to distinguish confounding from interaction. We computed Relative Excess Risks due to Interaction (RERI), Attributable Proportion (AP), and the Synergy index (S) to measure additive interaction. Results: High physical well-being is related to a large decrease in the risk of chronic pain (IRRTotal Effect = 0.58; 95% CI: 0.50-0.81). Perceived stress mediates 12.5% of the total effect of physical well-being on chronic pain. The stratum-specific IRRs of current smokers and non-current smokers were different from each other and were larger than the crude IRR (IRR = 1.49; 95% CI: 1.24-1.80), which indicates that smoking could be both confounder and interactor. Interaction analyses showed that physical activity could act as a potential interactor (RERI = 0.25; 95% CI: 0.13, 0.60). Conclusions: Perceived stress is an important mediator of the relation between physical well-being and chronic pain, while smoking is both a confounder and an interactor. Our findings may prove useful in distinguishing high-risk groups from low-risk groups, in the interventions aimed at reducing chronic pain.
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NK cells are the first sentinels of the immune system that can recognize and eradicate transformed cells. Their activation without a need for additional signaling have attracted great attention on the use of NK cells as a promising option in cancer immunotherapy. However, the large-scale production of NK cells for successful NK cells therapy is a challenge that needs to be tackled. Engineering NK cells to avoid tumor escape and improve their antitumor potency are the other matters of focus that have widely been studied in the recent years. This paper reviews the most recent advances in the stem cell-derived NK cell technology and discusses the potential of the engineered NK cells for clinical applications in cancer immunotherapy.
NK cells are important cells in our body's defense system that can find and destroy tumor cells. These cells are made in bone marrow (in adults) or umbilical cord (in the embryonic period) from a population of cells called stem cells, and then released into the blood and lymph. Stem cells are the early ancestral cells that can differentiate into multiple cell types. Because the number and function of NK cells in a tumor context are reduced, thus we can use these stem cells to make lots of NK cells for treatment purposes. Scientists can also make these cells even better at killing tumors by changing them to have special sensors. In the end, NK cells are like superheroes that fight and kill tumor cells, and using stem cells to make them is a really promising way to help treat malignant diseases.
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Células Matadoras Naturais , Neoplasias , Humanos , Imunoterapia , Imunoterapia Adotiva , Células-Tronco , Neoplasias/terapiaRESUMO
BACKGROUND: Chronic pain and depression represent two global health problems with considerable economic consequences. Although existing literature reports on the relation between depression and pain conditions, meta-analytic evidence backing the mediating role of sleep disturbance as one of the main symptoms of depression is scarce. To examine the extent to which sleep disturbance mediates the depression-chronic pain association, we conducted a systematic review and meta-analysis of the associations of chronic pain, depression, and sleep quality. METHODS: We systematically searched for literature in MEDLINE and other relevant databases and identified cohort and case-control studies on depression, sleep disturbance, and chronic pain. Forty-nine studies were eligible, with a total population of 120 489 individuals. We obtained direct and indirect path coefficients via two-stage meta-analytic structural equation modelling, examined heterogeneity via subgroup analyses, and evaluated primary studies quality. RESULTS: We found a significant, partial mediation effect of sleep disturbance on the relation between depression and chronic pain. The pooled path coefficient (coef.) of the indirect effect was 0.03 (95% confidence interval [CI]: 0.01-0.05) and accounted for 12.5% of the total effect of depression on chronic pain. This indirect effect also existed for cohort studies (coef. 0.02; 95% CI: 0.002-0.04), European studies (coef. 0.03; 95% CI: 0.004-0.05), and studies that adjusted for confounders (coef. 0.04; 95% CI: 0.01-0.09). CONCLUSIONS: Sleep disturbance partially mediates the association between depression and pain. Although plausible mechanisms could explain this mediation effect, other explanations, including reverse causation, must be further explored. SYSTEMATIC REVIEW PROTOCOL: PROSPERO CRD42022338201.
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Dor Crônica , Transtornos do Sono-Vigília , Humanos , Dor Crônica/complicações , Depressão/complicações , Qualidade do Sono , Sono , Transtornos do Sono-Vigília/complicações , Transtornos do Sono-Vigília/epidemiologiaRESUMO
Background: DNA probes have been widely used as diagnostic tools for translocations. This study sought to design a screening tool using ssDNA probes and chromosome conformation capture (3C) library fragment hybridization. Method: The authors focused on developing a probe for the juxtaposed region of MYC and TRD. Fragments of the MYC gene with a thiol modification (MYC-Au NP probe) were functionalized by gold nanoparticles (Au NPs). Then TRD probes were immobilized on a nitrocellulose surface. Hybridization between DNA probes and 3C library fragments of SKW3 cells was determined by color intensity. Results: Optimal hybridization of the 3C library sample of the cell line to probes showed higher color intensity than human umbilical vein endothelial cells. Conclusion: Combining 3C-based techniques and DNA-DNA hybridization can identify rearrangements in cancer cells.
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Técnicas Biossensoriais , Nanopartículas Metálicas , Humanos , Translocação Genética , Ouro , Células Endoteliais , Cromossomos , Sondas de DNA/genética , DNA/genética , Técnicas Biossensoriais/métodosRESUMO
BACKGROUND AND AIM: Adipocytokines play an important role in obstructive sleep apnea (OSA) by mediating inflammatory responses. Previous studies have reported that OSA is related to a change in the serum levels of adipocytokines; however, the results are still controversial. This meta-analysis aimed to assess the relationship between OSA and circulating level of adipocytokines in adults and children. METHODS: A comprehensive search was conducted in databases of Medline/PubMed and Scopus for pertinent articles published since their inception to July 2022. Weighted mean differences (WMDs) and 95% confidence intervals (CIs) were used to assess the strength of the relationship between the concentrations of adipocytokines with OSA. RESULTS: In the overall analysis, contrary to IL-10, which showed a significant reduction, IL-1ß, IL-4, IL-8, IL-17, and IFN- gamma showed higher levels in OSA patients in comparison with control groups (p <0.05). For adults, IL-1ß, IL-8, IL-17, IL-18, vaspin, visfatin, and chemerin were linked to a greater serum levels in patients with OSA, while, IL-5 and IL-10 were detected significantly lower in adults with OSA in comparison with healthy adults (p <0.05). In children with OSA, the serum levels of IL-4, IL-8, IL-12, IL-17, IL-23, and IFN-gamma were significantly higher than healthy children (p <0.05). CONCLUSION: The levels of inflammatory markers were found to be higher in OSA patients compared with control individuals, suggesting that adipocytokines may contribute to the pathology of OSA.
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Interleucina-10 , Apneia Obstrutiva do Sono , Criança , Adulto , Humanos , Interleucina-17 , Interleucina-8 , Adipocinas , Interleucina-4RESUMO
INTRODUCTION: Chronic pain represents a global health problem with a considerable economic burden. The relation of alcohol intake and chronic pain conditions was assessed in several studies with conflicting results. We used dose-response meta-analysis techniques to answer the question of whether alcohol intake is related to chronic pain occurrence. METHODS: We searched MEDLINE, Embase, and other databases to identify cohort and case-control studies on alcohol consumption and chronic pain. Sixteen studies were eligible with a total population of 642 587 individuals. Fixed-effects and random-effects pooled estimates were obtained by weighting log odds ratios (ORs) in case-control studies and log incidence rate ratios in cohort studies by the inverse of their variance. A heterogeneity assessment and a dose-response analysis were carried out. Quality scoring was also performed. RESULTS: Our results show that any alcohol consumption was related to lower odds of chronic pain (pooled OR=0.76; 95% confidence interval [CI], 0.61-0.95). The association was non-linear. The ORs by quartile of alcohol doses were as follows: OR2nd quartile=0.74; 95% CI, 0.64-0.87; OR3rd quartile=0.67; 95% CI, 0.53-0.86; and OR4th quartile=0.75; 95% CI, 0.50-1.14. This association was observed for cohort studies (OR=0.77; 95% CI, 0.61-0.98) and European studies (OR=0.65; 95% CI, 0.48-0.87) only. Studies with complete adjustment for confounding factors showed a stronger relation than those with incomplete adjustment (OR=0.69; 95% CI, 0.48-0.99 and OR=0.85; 95% CI, 0.65-1.11, respectively). CONCLUSION: Alcohol consumption presents a non-linear inverse association with the occurrence of chronic pain. Although plausible mechanisms could explain this protective effect, other explanations, including reverse causation, are probable.
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Dor Crônica , Consumo de Bebidas Alcoólicas/efeitos adversos , Consumo de Bebidas Alcoólicas/epidemiologia , Estudos de Casos e Controles , Dor Crônica/epidemiologia , Dor Crônica/etiologia , Etanol , Humanos , Fatores de RiscoRESUMO
Background: Despite the importance of self-esteem in cancer patients and the potential role of spiritual care programs in its promotion, no interventional study was found in this field in Iran. The aim of this study was therefore to investigate the effect of a spiritual care program on the self-esteem of patients with various types of cancer. Materials and Methods: This study was quasi-experimental with two groups using a before and after design which was conducted in 2019 in a selected hospital in Isfahan, Iran. A convenience sample of 64 patients randomly assigned into the experimental (n = 32) and the control group (n = 32). Data collection was conducted through a two-part questionnaire including Demographic characteristics and the Coopersmith Self-Esteem Inventory (SEI). The spiritual care program consisted of six sessions based on four domains including individuals' relationship with God, themselves, others, and with the environment. To analyze the data, Chi-square, Fisher's exact test, independent t test, and paired t test were performed. Results: The total self-esteem mean score and its domains were not significantly different between the two groups before the intervention (p > 0.05). However, after conducting the intervention, the total self-esteem mean score and its domains were significantly higher in the intervention group compared with the control group (p < 0.001). Conclusions: The spiritual care program used in this study was effective in promoting the self-esteem of patients with cancer. It is suggested that similar studies are conducted to provide fertile grounds for using such programs in the oncology clinical area.
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Type 1 diabetes is a metabolic disorder caused by the loss or dysfunction of ß-cells in the pancreas. Organ shortage is a critical concern of diabetic patients in need of beta islet transplantation. Tissue engineered islets are promising alternatives to traditional organ transplantation. Recent progress in stem cell biology and gene cloning techniques has raised hopes for the generation of insulin producing cells (IPCs) without the need of immunosuppression. The purpose of this study was to produce IPCs using human adipose-derived stem cells (hADSCs) and human endometrial-derived stem cells (hEnSCs) and also to compare the level of insulin secretion by these cells in 2D and 3D culture systems on fibrin scaffolding. Stem cells differentiation was carried out through transduction with an insulin over expression lentiviral vector. Real-time PCR and immunocytochemistry confirmed the successful transduction of both cell types. Both cell types showed comparable insulin secretion by ELISA.3D culture resulted in higher amounts of insulin secretion of the two cell types versus 2D as control. This study showed that insulin gene delivery to the stem cells could be an efficient method for producing IPCs and fibrin encapsulation enhances the functionality of these cells.
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Tecido Adiposo/metabolismo , Endométrio/metabolismo , Fibrina/química , Secreção de Insulina , Células-Tronco/metabolismo , Tecido Adiposo/citologia , Diferenciação Celular , Células Cultivadas , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/terapia , Endométrio/citologia , Feminino , Expressão Gênica , Terapia Genética/métodos , Células HEK293 , Humanos , Células Secretoras de Insulina , Transplante das Ilhotas Pancreáticas , Alicerces Teciduais , Transdução GenéticaRESUMO
The presence of biological cues to promote the attachment, proliferation, and differentiation of neuronal cells is important in the process of nerve regeneration. In this study, laminin as a neurite promoting protein, has been used to modify poly-lactide-co-glycolide/carbon nanotube (PLGA/CNT) electrospun nanofibrous scaffolds by means of either mussel-inspired poly(dopamine) (PD) coating or via direct physical adsorption as a simple route for the functionalization of biomaterials. The laminin-modified scaffolds were characterized by a combination of field emission scanning electron microscopy (SEM), X-ray photoelectron spectroscopy, and contact angle measurements. Subsequently, various properties of scaffolds such as degradation time, amount of attached laminin and the rate of CNT release were investigated. The synergistic effect of topographical and biological cues for PC12 cell attachment, proliferation, and differentiation were then studied by SEM and confocal microscopy. The results of degradation study showed that laminin-modified scaffolds were biodegradable with good structural integrity that persisted about 4 weeks. The amount of laminin attached to the PLGA/CNT and PLGA/CNT-PD scaffolds was 3.12 ± 0.6 and 3.04 ± 071 µg per mg of the scaffold, respectively. Although laminin-modified scaffolds could improve cell proliferation identically, neurite extensions on the PLGA/CNT scaffold modified via PD coating (PLGA/CNT-PD-lam scaffold) were significantly longer than those observed on PLGA/CNT scaffold modified via physical adsorption (PLGA/CNT-lam scaffold) and unmodified scaffolds. Together, these results indicated that surface modification via PD coating could be a promising strategy to fabricate biomimetic scaffolds capable of sustaining longer neuronal growth for nerve tissue engineering.
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Laminina/química , Nanotubos de Carbono/química , Tecido Nervoso , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Engenharia Tecidual , Alicerces Teciduais , Animais , Biomimética , Proliferação de Células/efeitos dos fármacos , Indóis/química , Microscopia Eletrônica de Varredura , Nanofibras/química , Células PC12 , Polímeros/química , Ratos , Propriedades de SuperfícieRESUMO
OBJECTIVES: This study aimed to assess the effect of photodynamic therapy (PDT) on expression of CASP3, NRAS and HRAS genes at mRNA levels, and apoptosis of head and neck squamous cell carcinoma (HNSCC) cell line. MATERIALS AND METHODS: In order to complete the present in vitro study, HNSCC cell line (NCBI C196 HN5) purchased from Pasteur Institute. Cells were divided into four groups; Group 1: photodynamic treatment (laser + methylene blue (MB) as photosensitizer), group 2: MB, group 3: laser (with 660 nm wavelength), and group 4: control (without any treatment). To determine the optimal concentration of MB, in a pilot study, toxicity of MB in different concentration was assessed using MTT assay. Cells in group 1, 2 and 3 was treated at optimal concentration of MB (1.6 µg/mL). Gene expression at mRNA levels was assessed after 24 h incubation, using real-time (qRT)-PCR. The expression of BAX and BCL2 genes at the mRNA levels was analyzed to evaluate apoptosis. 2-ΔΔCt values of BCL2, BAX, CASP3, NRAS, and HRAS in groups was analyzed using ANOVA. Tukey's HSD and Games Howell test was used to compare between two groups. RESULTS: Over-expression of BAX (p < 0.001), CASP3 (p < 0.001) and down-regulation of BCL2 (p = 0.004), HRAS (p = 0.023) and NRAS (p = 0.045) were noted in group 1 (PDT), compared with the control group. Treatment by laser alone induce down-regulation of CASP3 (p < 0.05), BAX (p < 0.05), BCL2 (p > 0.05), HRAS (p > 0.05) and NRAS (p > 0.05). CONCLUSION: PDT caused down-regulation of NRAS, HRAS and BCL2 and over-expression of CASP3 and BAX genes at mRNA levels in HNSCC cell line. The present study raises the possibility that the role of MB on BCL2 down-regulation and BAX and CASP3 over-expression was higher than laser alone while it seems that laser alone was more effective than MB in HRAS and NRAS down-regulation.
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Neoplasias de Cabeça e Pescoço , Fotoquimioterapia , Apoptose , Caspase 3 , Linhagem Celular , Linhagem Celular Tumoral , GTP Fosfo-Hidrolases/genética , GTP Fosfo-Hidrolases/farmacologia , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Neoplasias de Cabeça e Pescoço/genética , Humanos , Proteínas de Membrana/genética , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Projetos Piloto , Proto-Oncogene Mas , Proteínas Proto-Oncogênicas p21(ras)/genética , Proteínas Proto-Oncogênicas p21(ras)/farmacologia , RNA Mensageiro , Carcinoma de Células Escamosas de Cabeça e Pescoço/tratamento farmacológico , Carcinoma de Células Escamosas de Cabeça e Pescoço/genéticaRESUMO
Background: Chromosomal breakpoints are the most common cause of hereditary diseases and cancers. Today, many standard clinical methods such as cytogenetic and PCR based techniques are used which have limitation regarding detection resolution. Chromosome conformation capture is a method for detecting gene proximity and chromosomal rearrangements. Materials and Methods: In this study, SKW3 cell line was used for detecting t(8;14)(q24;q11) using a 3C-based technique. SKW3 cell line was used for 3C library preparation. For Inverse PCR, two regions were selected in upstream and downstream of the viewpoint locus on chromosome 8-MYC gene based on EcoRI restriction sites. The captured sequence with intra-chromosomal interaction between chr8-c-MYC and chr14-TRD was selected for the translocation PCR primer design. Results: The DNA fragment captured in 3C PCR showed a specific TRD sequence translocated downstream of the MYC gene. Translocation PCR demonstrated the existence of (8; 14) (q24; q11) MYC /TRD in both library and genomic DNA. Conclusion: This result demonstrated 3C- based method could be used as a useful low-cost easy operating technique in chromosomal rearrangements detection. In this study, the integration of whole genome library monitoring and PCR method was used as a high- through put method in chromosomal breakpoints detection.
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In the current study, electrospun poly(ε-caprolactone)-gelatin (PCL-Gel) fibrous scaffolds containing magnesium oxide (MgO) particles and preseeded with human endometrial stem cells (hEnSCs) were developed to use as wound care material in skin tissue engineering applications. Electrospun fibers were fabricated using PCL-Gel (1:1 [wt/wt]) with different concentrations of MgO particles (1, 2, and 4 wt%). The fibrous scaffolds were evaluated regarding their microstructure, mechanical properties, surface wettability, and in vitro and in vivo performances. The full-thickness excisional wound model was used to evaluate the in vivo wound healing ability of the fabricated scaffolds. Our findings confirmed that the wounds covered with PCL-Gel fibrous scaffolds containing 2 wt% MgO and preseeded with hEnSCs have nearly 79% wound closure ability while sterile gauze showed 11% of wound size reduction. Our results can be employed for biomaterials aimed at the healing of full-thickness skin wounds.
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Endométrio/metabolismo , Gelatina/química , Óxido de Magnésio/química , Poliésteres/química , Pele/metabolismo , Células-Tronco/metabolismo , Alicerces Teciduais/química , Cicatrização , Ferimentos e Lesões , Animais , Feminino , Xenoenxertos , Humanos , Masculino , Porosidade , Ratos , Ratos Wistar , Transplante de Células-Tronco , Ferimentos e Lesões/metabolismo , Ferimentos e Lesões/terapiaRESUMO
BACKGROUND: Astragalus fascicolifolius manna is used to treat different diseases. Because pregnant women tend to use Astragalus. fascicolifolius and Iranian traditional medicine emphasizes the abortifacient potential of this plant, this study aimed to investigate Astragalus fascicolifolius manna abortifacient property and effects on estrogen, progesterone, LH and FSH levels in BALB/c mice. METHOD: This experimental study was conducted with 70 female BALB/c mice assigned to seven groups: Nonpregnant, untreated; nonpregnant, Astragalus. fascicolifolius extract (400 mg/kg)-treated; pregnant, Astragalus. fascicolifolius extract (400, 800 and 1200 mg/kg)-treated; and two pregnant control groups. On 18 and 19 days of pregnancy, cesarean section performed on mice, resorbed embryos counted; then Follicle-stimulating hormone (FSH), Luteinizing hormone (LH), estrogen and progesterone levels were measured by the ELISA. RESULTS: Astragalus. fascicolifolius extract caused a significant increase abortion in mice. The levels of progesterone, FSH and LH were significantly different among the groups such that mean progesterone level was lower and mean LH and FSH levels were higher in the Astragalus. fascicolifolius extract-treated groups than the pregnant, untreated group. CONCLUSION: This extract has abortifacient properties and this plant can be used cautiously in pregnancy. Decreasing progesterone, increasing FSH and LH feedback in response to decreased progesterone by this extract is one of the potential mechanisms involved in abortion.
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â¢It is good to know that sensitivity is an important measure in public health aspects instead of clinical fields. Likewise, PPV and NPV are among the measures which are more appropriate for advice about the validity of a diagnostic test for clinical purposes.â¢It is critically important to consider that accurate diagnosis is very important in appendicitis because of the urgent need for surgery in people who are truly positive, likewise, operating surgery for people who are not truly positive will have inconvenient effects.â¢Finally, for the prediction of an outcome, we need data from two different cohorts or at least from one cohort divided into two to first to develop a prediction model and subsequently validate it.â¢To make our methodological comments brief, it is crucial to know that to determine the validity and predictive ability of ultrasound in predicting acute appendicitis among children, methodological and statistical issues should be correctly taken into account.