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BACKGROUND: Pulmonary fibrosis (PF) leads to excessive deposition of fibrous connective tissue in the lungs, increasing the risk of lung cancer due to the enhanced activity of fibroblasts (FBs). Fibroblast-mediated collagen fiber deposition creates a tumor-like microenvironment, laying the foundation for tumorigenesis. Clinically, numerous cases of lung cancer induced by pulmonary fibrosis have been observed. In recent years, the study of nucleotide point mutations, which provide more detailed insights than gene expression, has made significant advancements, offering new perspectives for clinical research. METHODS: We initially employed Mendelian randomization to ascertain that the initial stage of lung cancer induced by PF belongs to small cell lung cancer (SCLC). Subsequently, pulmonary neuroendocrine cells (PNECs) were identified by using pseudo-time series analysis as cell clusters with carcinogenic potential. We categorized FBs into four groups according to their cellular metabolism, and then analyzed the cellular communication between FBs and PNECs, as well as changes in intracellular pathways of PNECs. Additionally, we examined the characteristic genome of FBs which is significantly associated with PF and investigated the impact of FBs on immune cells in the PF microenvironment. Finally, we explored strategies for preventing the progression from PF to lung cancer. RESULTS: The genetic features of cells with carcinogenic potential in PF tissues were revealed, characterized by upregulation of Achaete-Scute Family BHLH Transcription Factor 1 (ASCL1), Homeobox B2 (HOXB2), Teashirt Zinc Finger Homeobox 2 (TSHZ2), Insulinoma-associated 1 (INSM1), and reduced activity of RE1 Silencing Transcription Factor (REST). FBs characterized by high glycolysis and low tricarboxylic acid (TCA) cycling played a key role in the progression of PF. The microenvironment of PF resembles the tumor microenvironment, providing a conducive immunosuppressive environment for the occurrence of cancer cells. In dendritic cells, rs9265808 is a susceptibility locus for progression from pulmonary fibrosis to lung cancer, mutations at this locus increase the expression of Complement Factor B (CFB), and excessive activation of the complement pathway is a crucial factor leading to lung cancer development in patients with pulmonary fibrosis. Ensuring adequate nutritional supply and physical function is one of the effective measures to prevent progression from pulmonary fibrosis to lung cancer. CONCLUSION: CFB promotes lung cancer occurrence by inducing the accumulation and polarization of a large number of monocytes/macrophages in the lungs, driving disease progression by reducing the physical fitness of patients with pulmonary fibrosis.
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BACKGROUND: This systematic review and meta-analysis aimed to evaluate the effects of Taohong Siwu Decoction (THSWD) combined with low molecular weight heparin (LMWH), as well as THSWD alone, on the incidence of Deep vein thrombosis (DVT), D-dimer levels, prothrombin time (PT), activated partial thromboplastin time (APTT), visual analogue scale (VAS) pain score, and calf swelling in patients undergoing hip fracture or replacement surgery, compared to LMWH. METHODS: According to the predefined inclusion criteria, we conducted a comprehensive search for randomized controlled trials (RCTs) examining the efficacy of THSWD combined with LMWH or THSWD compared to LMWH in patients with hip fractures or undergoing replacement surgery. The search was performed across multiple databases including China National Knowledge Internet, WanFang, Sinomed, Duxiu, PubMed, Embase, Google Scholar, Cochrane, and Web of Science from their inception until December 2023. Additionally, relevant literature references were retrieved and hand searching of pertinent journals was conducted. The methodological quality assessment of the included trials was carried out following the guidelines outlined in the Cochrane Handbook. Review Manager 5.4 was applied in analyzing and synthesizing. RESULTS: A total of 18 RCTs with 1353 patients were included. The results of meta-analysis showed that compared with the control group, the combined group had a better effect on the incidence of DVT [RRâ =â 0.32, 95% CI(0.17, 0.58; Pâ =â .0002], D-dimer [SMDâ =â -5.88, 95% CI(-7.66, -4.11); Pâ <â .00001], VAS [MDâ =â -1.16, 95% CI(-1.81, -0.50); Pâ =â .0005], Calf circumference difference [MDâ =â -0.56, 95% CI(-1.05, -0.08); Pâ =â .02]. There was no significant difference in PT and APTT between the combined group and the control group. Meta-analysis results show that the D-dimer, incidence of DVT, PT, and APTT did not significantly differ between the THSWD and the LMWH groups. CONCLUSION: This meta-analysis shows that compared with LMWH, THSWD combined with LMWH has a better efficacy in the treatment of DVT after hip surgery, without a significant increase in the incidence of adverse events. Additionally, the combined therapy can also reduce D-dimer, VAS, and swelling. However, due to the limitations of the included studies (such as small sample size and low-quality evidence), the results need to be further verified in more rigorous multicenter clinical trials with a large sample size.
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Anticoagulantes , Medicamentos Herbarios Chinos , Trombosis de la Vena , Humanos , Anticoagulantes/uso terapéutico , Ensayos Clínicos Controlados Aleatorios como Asunto , Heparina de Bajo-Peso-Molecular , Trombosis de la Vena/tratamiento farmacológico , Estudios Multicéntricos como AsuntoRESUMEN
INTRODUCTION: Lung cancer is the primary cause of cancer-related deaths worldwide, with high rates of morbidity and mortality. The most effective treatment for early stage (I-II) non-small cell lung cancer (NSCLC) is surgical resection. However, the extent of mediastinal lymph nodes removal required and the impact of their removal remains controversial. This systematic review and meta-analysis aimed to evaluate the postoperative complications in patients with stage I-II NSCLC who received mediastinal lymph node dissection (MLND) or mediastinal lymph node sampling (MLNS). METHODS AND ANALYSIS: According to the predefined inclusion criteria, we will conduct a comprehensive search for randomized controlled trials (RCTs) and observational studies examining the postoperative complications of MLND compared to MLNS in patients with stage I-II NSCLC. The search will be performed across multiple databases including PubMed, Embase, the Cochrane Library, CNKI, WanFang, Sinomed, VIP, Duxiu, and Web of Science from inception to February 2024. Additionally, relevant literature references will be retrieved and hand searching of pertinent journals will be conducted. Screening, data extraction, and quality assessment will be performed by two independent reviewers. Review Manager 5.4 will be applied in analyzing and synthesizing. The Grading of Recommendations Assessment, Development and Evaluation (GRADE) will be used to assess the quality of evidence for the whole RCTs and used Newcastle-Ottawa scale to assess the methodologic quality of observational studies. ETHICS AND DISSEMINATION: This study did not include personal information. Ethical approval was not required for this study. This study is based on a secondary analysis of the literature, so ethical review approval is not required. The final report will be published in a peer-reviewed journal. CONCLUSION: This systematic review will contribute to compare the safety and survival benefits of these two surgical techniques for the treatment of early stage NSCLC, to further guide the selection of surgical approaches. TRIAL REGISTRATION: The protocol of the systematic review has been registered on Open Science Framework, with a registration number of DOI https://doi.org/10.17605/OSF.IO/N2Y5D.
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Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Escisión del Ganglio Linfático , Mediastino , Metaanálisis como Asunto , Complicaciones Posoperatorias , Revisiones Sistemáticas como Asunto , Humanos , Carcinoma de Pulmón de Células no Pequeñas/cirugía , Carcinoma de Pulmón de Células no Pequeñas/patología , Escisión del Ganglio Linfático/efectos adversos , Escisión del Ganglio Linfático/métodos , Neoplasias Pulmonares/cirugía , Neoplasias Pulmonares/patología , Complicaciones Posoperatorias/etiología , Mediastino/patología , Mediastino/cirugía , Ganglios Linfáticos/patología , Ganglios Linfáticos/cirugía , Estadificación de NeoplasiasRESUMEN
With the improvement of global dietary conditions, non-alcoholic fatty liver disease (NAFLD) has gradually become prevalent. As the number of NAFLD patients increases, the coexistence of diseases associated with it has come into focus. In this study, based on immune phenotypes, intercellular communication activities, and clinical manifestations of NAFLD patients, IL1RN was identified as a central pro-inflammatory factor. Subsequently, potential downstream biological pathways of IL1RN in liver tissues and various cell types were enriched to describe its functions. Transcription factors Nfkb1, Jun, and Sp1, significantly associated with these functions, were also enriched. Functional studies of IL1RN suggest its potential to trigger autoimmune diseases. Given this, Mendelian randomization analysis was used to explore the causal relationship between NAFLD and various autoimmune diseases, with IL1RN considered as an intermediary introduced into Mendelian randomization studies. The results indicate that IL1RN and its partially related proteins play a certain mediating role in the process of NAFLD inducing rheumatoid arthritis (RA). Finally, additional research results suggest that intrahepatic ALT levels may influence IL1RN levels, possibly through amino acid metabolism.
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Artritis Reumatoide , Enfermedades Autoinmunes , Enfermedad del Hígado Graso no Alcohólico , Humanos , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Artritis Reumatoide/genética , Fenotipo , Estudio de Asociación del Genoma Completo , Proteína Antagonista del Receptor de Interleucina 1/genéticaRESUMEN
In both cancer and infections, diseased cells are presented to human Vγ9Vδ2 T cells through an 'inside out' signalling process whereby structurally diverse phosphoantigen (pAg) molecules are sensed by the intracellular domain of butyrophilin BTN3A11-4. Here we show how-in both humans and alpaca-multiple pAgs function as 'molecular glues' to promote heteromeric association between the intracellular domains of BTN3A1 and the structurally similar butyrophilin BTN2A1. X-ray crystallography studies visualized that engagement of BTN3A1 with pAgs forms a composite interface for direct binding to BTN2A1, with various pAg molecules each positioned at the centre of the interface and gluing the butyrophilins with distinct affinities. Our structural insights guided mutagenesis experiments that led to disruption of the intracellular BTN3A1-BTN2A1 association, abolishing pAg-mediated Vγ9Vδ2 T cell activation. Analyses using structure-based molecular-dynamics simulations, 19F-NMR investigations, chimeric receptor engineering and direct measurement of intercellular binding force revealed how pAg-mediated BTN2A1 association drives BTN3A1 intracellular fluctuations outwards in a thermodynamically favourable manner, thereby enabling BTN3A1 to push off from the BTN2A1 ectodomain to initiate T cell receptor-mediated γδ T cell activation. Practically, we harnessed the molecular-glue model for immunotherapeutics design, demonstrating chemical principles for developing both small-molecule activators and inhibitors of human γδ T cell function.
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Butirofilinas , Activación de Linfocitos , Fosfoproteínas , Receptores de Antígenos de Linfocitos T gamma-delta , Linfocitos T , Animales , Humanos , Antígenos CD/inmunología , Antígenos CD/metabolismo , Butirofilinas/inmunología , Butirofilinas/metabolismo , Camélidos del Nuevo Mundo/inmunología , Simulación de Dinámica Molecular , Fosfoproteínas/inmunología , Fosfoproteínas/metabolismo , Receptores de Antígenos de Linfocitos T gamma-delta/inmunología , Receptores de Antígenos de Linfocitos T gamma-delta/metabolismo , Linfocitos T/citología , Linfocitos T/inmunología , Linfocitos T/metabolismo , Cristalografía por Rayos X , Resonancia Magnética Nuclear Biomolecular , TermodinámicaRESUMEN
Ozone (O3) has become the most critical air pollutant in the Yangtze River Delta (YRD) region of China. Research on the O3 formation mechanism and its precursor sources (including nitrogen oxides (NOX) and volatile organic compounds (VOCs)) could provide a theoretical basis for mitigating O3 pollution in this region. In this study, simultaneous field experiments were conducted for air pollutants in a typical urban area (Suzhou) in the YRD region in 2022. The capacity of in-situ O3 formation, O3-NOX-VOCs sensitivities and sources of O3 precursors were analyzed. The results showed that in-situ formation contributed 20.8 % of the O3 concentration in the warm season (April to October) of the Suzhou urban area. Compared with the warm season average, the concentrations of various O3 precursors increased on pollution days. The O3-NOX-VOCs sensitivity was the VOCs-limited regime based on the average concentrations during the warm season. O3 formation was most sensitive to anthropogenic VOCs, of which oxygenated VOCs, alkenes and aromatics were the key species. There was a VOCs-limited regime in spring and autumn, while a transitional regime in summer due to the changes in NOX concentrations. This study considered NOX emission from VOCs sources and calculated the contribution of various sources to O3 formation. The results of VOCs source apportionment showed that diesel engine exhaust and fossil fuel combustion had a dominant proportion, but O3 formation presented significant negative sensitivities to the above two sources because of their high NOX emissions. There were significant sensitivities of O3 formation to gasoline vehicle exhaust and VOCs evaporative emissions (gasoline evaporation and solvent usage). The contribution of VOCs evaporative emissions during the O3 pollution episode was significantly higher than the average; therefore, controlling VOCs evaporative emissions during the O3 pollution episode is critical. These results provide feasible strategies to mitigate O3 pollution.
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Contaminantes Atmosféricos , Ozono , Compuestos Orgánicos Volátiles , Ozono/análisis , Gasolina , Monitoreo del Ambiente/métodos , China , Contaminantes Atmosféricos/análisis , Compuestos Orgánicos Volátiles/análisis , Emisiones de Vehículos/análisisRESUMEN
Feeder cells are essential to derive pluripotent stem cells (PSCs). Mouse embryonic fibroblasts (MEF) are widely used as feeder to generate and culture embryonic stem cells (ESCs) and induced PSCs (iPSCs) in many species. However it may not be suitable for livestock ESCs/iPSCs due to interspecies difference. Previously we derived bovine iPSCs from bovine Sertoli cells using MEF feeder. Here we compared the effects of MEF feeder and bovine embryonic fibroblasts (BEF) feeder on the maintenance of bovine iPSC pluripotency and morphology as well their contributions to the naïve-like conversion, based on a naïve medium (NM). The results showed successful conversion of the primed bovine iPSCs to naïve-like state within 3-4 days both on MEF feeder and BEF feeder in NM (termed as MNM and BNM respectively). These naïve-like iPSCs showed normal karyotype. There were more iPSC colonies under BNM condition than MNM condition. Epigenetically, histone modification H3K4 was upregulated, while H3K27 was downregulated in the naïve-like iPSCs. We further analyzed the naïve markers and differentiation potential both in vitro and in vivo of these cells, which were all reserved throughout the maintenance. Together, bovine naïve-like iPSCs can be generated both on MEF and BEF feeder in NM condition. The BNM condition is able to sustain the pluripotency and differentiation potential of the naïve-like bovine iPSCs, and improve the conversion efficiency.
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Células Madre Pluripotentes Inducidas , Ratones , Animales , Bovinos , Masculino , Células de Sertoli , FibroblastosRESUMEN
Schisandrin A and B (Sch A and B) are the important components of Asian dietary supplement and phytomedicine Schisandra chinensis (S. chinensis). They can enhance adult neurogenesis in vivo; however, these effects still need to be verified. Here NE-4 C neural stem cells (NSCs) were employed as the in vitro model and treated with Sch A and B at 0.1 µg/mL. EdU (5-Ethynyl-2'-deoxyuridine) labeling showed that both Sch A and B treatments enhanced NSC proliferation. Real-time PCR analysis showed the mRNA abundances of telomerase gene Tert and cell cycle gene Cyclin D1 were significantly up-regulated after the treatments. During the neurosphere induction, Sch B enhanced the neurosphere formation and neuronal differentiation, and increased the neurosphere semidiameters. Detection of the neuron differentiation marker Mapt indicates that both Sch A and B, especially Sch B, benefits the induced neuronal differentiation. Sch B treatment also enhanced mRNA expressions of the neurosphere-specific adhesion molecule Cdh2 and Wnt pathway-related genes including Mmp9, Cyclin D1 and ß-catenin. Together, Sch A especially Sch B, promotes the proliferation, affects the survival, differentiation and neurogenesis of NSCs, which is consistent with their in vivo effects. This study provides further clue on the potential neuropharmacological effects of S. chinensis.
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Células-Madre Neurales , Diferenciación Celular , Proliferación Celular , Ciclooctanos , Lignanos , Neurogénesis , Compuestos PolicíclicosRESUMEN
To study the carbon components in indoor and outdoor PM2.5, the samples of PM2.5 were collected from Nankai University in December 2015. The contents of eight carbon components were analyzed to use the thermo-optical reflection method. The results indicated that organic carbon (OC) mass concentration was 17.01, 19.48 and 18.92 µg/m3 in outdoor, dormitory and laboratory; elemental carbon (EC) mass concentration was 7.97, 3.56 and 3.53 µg/m3 in outdoor, dormitory and laboratory; and the total carbon aerosol was the proportion of more than 23% of PM2.5 samples. Lower wind speed and higher relative humidity were helpful to the accumulation of PM2.5. The ratio of OC/EC was > 2, and the SOC/OC ratio was > 30%, indicating that SOC was a crucial component indoors and outdoors. About 72% and 85% of the outdoor OC entering dormitory and laboratory environment, and about 59% and 71% of the outdoor EC entering dormitory and laboratory environment. Factor analysis of the eight carbon fractions indicated that the sources of OC and EC in outdoor, dormitory and laboratory is different.
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OBJECTIVE: Sertoli cells (SCs) are important sustentacular cells in the seminiferous tubules of the testis. Isolation and identification of SCs are the premise for studying their functions. Since New Zealand rabbit is a stable strain which is widely used for biomedical research and animal farming, this study aimed to develop a simple and effective protocol for SC isolation in New Zealand rabbits. MATERIALS AND METHODS: The SCs of three 30-day-old New Zealand rabbits were isolated by incubation with enzymatic digestion I (Dulbecco's modified Eagle medium supplemented with 1 mg/ml collagenase IV and 50 µg/ml DNase I) and digestion II (digestion I + 1 mg/ml hyaluronidase + 1 mg/ml trypsin), as well as differential plating. The cells were enriched and identified by using immunocytochemical staining and reverse transcription polymerase chain reaction analysis. RESULTS: Homogeneous cells were obtained. They presented the typical large cell body and an irregular pyramidal shape after differential plating and passaging. These cells expressed mRNA of the SC marker sex-determining region Y-box 9 (SOX9) instead of the Leydig cell marker StAR. Immunocytochemically, they are positive of SOX9, GATA binding protein 4, and androgen-binding protein. CONCLUSION: The SCs were enriched from the testicular tissues of prepubertal New Zealand rabbits by a simple and effective protocol, which provides a basis for further theoretical researches and practical applications.
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Germplasm preservation of livestock or endangered animals and expansion of germline stem cells are important. The purpose of this study is to investigate whether supplementation of trehalose to the freezing medium (FM) reduces tissular damage and improves the quality of testicular cells in the cryopreserved bovine testicular tissues. We herein established an optimized protocol for the cryopreservation of bovine testicular tissues, and the isolation as well as culture of bovine germ cells containing spermatogonial stem cells (SSCs) from these tissues. The results showed that FM containing 10% dimethyl sulfoxide (Me2SO/DMSO), 10% knockout serum replacement (KSR) and 20% trehalose (FM5) combined with the uncontrolled slow freezing (USF) procedures has the optimized cryoprotective effect on bovine testicular tissues. The FM5 + USF protocol reduced the cell apoptosis, maintained high cell viability, supported the structural integrity and seminiferous epithelial cohesion similar to that in the fresh tissues. Viable germ cells containing SSCs were effectively isolated from these tissues and they maintained germline marker expressions in the co-testicular cells and co-mouse embryonic fibroblasts (MEF) feeder culture systems respectively, during the short-term culture. Additionally, upregulated transcriptions of spermatogenic differentiation marker C-KIT and meiotic marker SYCP3 were detected in these cells after retinoic acid-induced differentiation. Together, FM5 + USF is suitable for the cryopreservation of bovine testicular tissues, with benefits of reducing the apoptosis, maintaining the cell viability, supporting the testicular structure integrity, and sustaining the survival and differentiation potential of bovine germ cells containing SSCs.
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Criopreservación , Trehalosa , Animales , Bovinos , Supervivencia Celular , Criopreservación/métodos , Crioprotectores/farmacología , Dimetilsulfóxido , Fibroblastos , Masculino , Ratones , Espermatogonias , Testículo , Trehalosa/farmacologíaRESUMEN
Germplasm cryopreservation and expansion of gonocytes/prospermatogonia or spermatogonial stem cells (SSCs) are important; however, it's difficult in cattle. Since inhibitors of Mek1/2 and Gsk3ß (2i) can enhance pluripotency maintenance, effects of 2i-based medium on the cultivation of bovine prospermatogonia from the cryopreserved tissues were examined. The testicular tissues of newborn bulls were well cryopreserved. High mRNA levels of prospermatogonium/SSC markers (PLZF, GFRα-1) and pluripotency markers (Oct4/Pouf5, Sox2, Nanog) were detected and the PLZF+ /GFRα-1+ prospermatogonia were consistently identified immunohistochemically in the seminiferous cords. Using differential plating and Percoll-based centrifugation, 41.59% prospermatogonia were enriched and they proliferated robustly in 2i medium. The 2i medium boosted mRNA abundances of Pouf5, Sox2, Nanog, GFRα-1, PLZF, anti-apoptosis gene Bcl2, LIF receptor gene LIFR and enhanced PLZF protein expression, but suppressed mRNA expressions of spermatogonial differentiation marker c-kit and pro-apoptotic gene Bax, in the cultured prospermatogonia. It also alleviated H2 O2 -induced apoptosis of the enriched cells and decreased histone H3 lysine (K9) trimethylation (H3K9me3) and its methylase Suv39h1/2 mRNA level in the cultured seminiferous cords. Overall, 2i medium improves the cultivation of bovine prospermatogonia isolated from the cryopreserved testes, by inhibiting Suv39h1/2-mediated H3K9me3 through Mek1/2 and Gsk3ß signalling, evidencing successful cryopreservation and expansion of bovine germplasm.
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Células Madre Germinales Adultas , Espermatogonias , Animales , Bovinos , Criopreservación , Medios de Cultivo , Masculino , TestículoRESUMEN
Silicon carbide (SiC) ceramic is an ideal material for mechanical seal because of its super hardness, high strength, low friction coefficient, good thermal conductivity, and resistance to friction and wear. However, due to relatively high resistivity of SiC ceramic, the triboelectric charge caused by rubbing of mechanical seal end-faces could not be released. It is terrible that the accumulation of triboelectric charge could cause electrochemical corrosion, which would accelerate wear. To decrease the resistivity of SiC ceramic is a desire for improving the performance of mechanical seal. In this research, decreasing resistivity of pressureless sintered SiC ceramic was investigated by conductive pathways through semiconductive grains in a body by incorporation of graphene, which has an extremely low resistivity. With the increasing of graphene from 0 to 2 wt.%, the volume resistivity of SiC ceramics sintered with graphene decreased logarithmically from >106 to around 200 Ω·cm, and the bulk density decreased gradually, from 3.132 to 3.039 g/cm3. In order to meet the requirements of mechanical seal, SiC ceramic sintered with 1 wt.% of graphene, for which the volume resistivity is of 397 Ω·cm, the bulk density is of 3.076 g/cm3, and the flexural strength is of 364 MPa, was optimized when all properties were taken into consideration. It is possible to fabricate low-resistivity SiC ceramic as a useful friction pair material for mechanical seal in a special condition, without excessive loss of their excellent mechanical properties by the introduction of partially connected graphene as conductive pathway into semiconducting ceramic.
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Sertoli cells (SC) have important functions in spermatogenesis by regulating development of spermatogenic cells. Glial cell line-derived neurotrophic factor (GDNF) are produced by SC. Although the effects of GDNF on spermatogenesis have been well studied, the understanding of how GDNF is synthesized is still limited, especially in food animal producing species. Because protein kinase (PK) has varied functions in multiple cellular processes and the PK pathway modulates SC functions, the objective of the present study was to determine whether PK modulates the abundance of GDNF protein in SC of cattle. To conduct this study, immature SC were enriched from cryopreserved testicular tissues of 1-day-old bulls. These cells had a marked proliferation capacity. Results from immunostaining analysis indicated that there was a sustained abundance of SC mRNA marker protein transcripts and marker proteins: androgen bind protein (ABP), GATA4 and VIMENTIN. There was subsequent characterization of SC treated with the PK inhibitor staurosporine for 0, 1 or 2 h. Results from real-time-PCR and Western blot analyses indicated the treatment (2 h) resulted in a decrease in Gdnf mRNA transcript and GDNF protein. Additionally, the staurosporine treatment resulted in an increase in the abundance of anti-apoptosis Bcl2 and decrease in pro-apoptosis Bax mRNA transcripts. Furthermore, results of the TUNEL assay indicated there was a decrease in apoptosis in the staurosprine-treated SC. Collectively, results indicate the PK signaling is involved in regulation of GDNF protein abundance in the immature SC and the survival of these cells in cattle.
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Regulación de la Expresión Génica/efectos de los fármacos , Factor Neurotrófico Derivado de la Línea Celular Glial/metabolismo , ARN Mensajero/metabolismo , Células de Sertoli/metabolismo , Estaurosporina/farmacología , Animales , Animales Recién Nacidos , Bovinos , Factor Neurotrófico Derivado de la Línea Celular Glial/genética , Masculino , ARN Mensajero/genética , Maduración SexualRESUMEN
Schisandrin A and B (Sch A and B) are the main effective components of Schisandra chinensis (S. chinensis), which is traditionally used to enhance mental and intellectual functions in eastern Asia. Previously, we reported Sch A and B remarkably affect adult neurogenesis in the subventricular zone of mouse lateral ventricle. Since the neurogenesis in the hippocampal dentate gyrus (DG) is more important to learning, memory and cognition, here we further examined their effects on the adult DG neurogenesis. Phosphohistone H3 (PHH3) immunostaining showed that Sch B significantly enhanced the cell proliferation in the DG. Glial fibrillary acidic protein (GFAP, mostly labels astrocytes and some stem cells) staining was used to further identify the proliferating cell type. Dramatically, increases of GFAP+ cells in both Sch A and B treated groups were observed. What's more, the total numbers of the mature neurons labeled by neuron-specific nuclear protein (NeuN) were also increased in both Sch A and B treated groups compared with the controls. Together, Sch A and B enhance the adult DG neurogenesis by increasing astrocytes/stem cells and improving the survival and maturation of DG neurons. Our study shed a new light on the neuropharmacological functions of the herbal medicine S. chinensis.
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Ciclooctanos/farmacología , Giro Dentado/efectos de los fármacos , Lignanos/farmacología , Neurogénesis/efectos de los fármacos , Neuronas/efectos de los fármacos , Compuestos Policíclicos/farmacología , Animales , Proliferación Celular/efectos de los fármacos , Hipocampo/efectos de los fármacos , Masculino , RatonesRESUMEN
To enrich bovine gonocytes from cryopreserved testicular tissues, the cryoprotection effects of the freezing media containing knockout serum replacement (KSR) were examined. Using Minimum essential medium (MEM) + 10% dimethyl sulfoxide (Me2SO) as the basic medium, calf testicular tissues were cryopreserved in media containing 0, 5, 10, 20, 40, 90% KSR and 5% fetal bovine serum (FBS) respectively. Morphologically, the seminiferous cords and interstitium were well preserved in all groups. The gonocytes were all glial cell line-derived neurotrophic factor (GDNF) family receptor α-1 (GFRα-1) positive. The recovery rates in all KSR groups were higher than that of the 10% Me2SO group, while comparable to the 5% FBS group. The enriched gonocytes expressed gonocyte marker GFRα-1 typically. Collectively, supplementation of 5-10% KSR can achieve comparable cryoprotective effects with using 5% FBS, which is useful in future study due to its defined formulation that is more consistent in quality and stable in supply.
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Criopreservación/métodos , Crioprotectores/farmacología , Suero/metabolismo , Testículo/citología , Animales , Bovinos , Supervivencia Celular/efectos de los fármacos , Dimetilsulfóxido/farmacología , Congelación , Receptores del Factor Neurotrófico Derivado de la Línea Celular Glial/metabolismo , Masculino , Suero/químicaRESUMEN
Although induced pluripotent stem cells (iPSCs) had been generated from several somatic cell types in cattle, their pluripotency and differentiation capacities after freezing/thawing, and the dysregulated transcripts involved in pathways critical for reprogramming were not investigated. Additionally, selection of proper source cells is critical for iPSC derivation because the residual influence of the somatic origin may variegate their differentiation propensity. Sertoli cells (SCs) have special properties suitable for iPSCs derivation. Herein bovine SCs were enriched from the cryopreserved testicular tissues and reprogrammed into iPSCs using lentivirus carrying yamanaka factors (OSKM). These iPSCs have typical morphology resembling human iPSCs and remain normal karyotypes. They can express alkaline phosphatase activity and common pluripotency markers with a low methylation in the promoter region of Nanog. They can also form embryoid bodies and teratomas that give rise to cells/tissues from three embryonic germ layers. Transcriptome profiling showed that the exogenous OSKM were silenced and 8009 dysregulated mRNAs were identified. The pluripotency, methyldioxygenase and anti-apoptosis genes were all upregulated but the apoptotic gene downregulated in these iPSCs. Bunch of pathways related to the reprogramming, inflammation and viral infection pathways were upregulated, while pathways associated with the differentiation, senescence, metabolism and apoptosis were downregulated in these cells. After cryopreservation/thawing, the recovered iPSCs remain strong pluripotency and differentiation capabilities. Together, iPSCs were derived from the bovine SCs isolated from the cryopreserved neonatal bull testis, pluripotency and differentiation capacities verified, iPSCs cryopreserved, cultured and again reverified for pluripotency and differentiation capacities.
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Bovinos , Células Madre Pluripotentes Inducidas/fisiología , Células de Sertoli/fisiología , Transcriptoma , Animales , Reprogramación Celular , Criopreservación/veterinaria , Cuerpos Embrioides , Regulación de la Expresión Génica , MasculinoRESUMEN
The direct ligands recognized by γδ T cell receptors (γδ TCRs) remain uncertain and controversial. In a study appearing in this issue, Willcox et al. use surface plasmon resonance and isothermal titration calorimetry to demonstrate that B7-like molecule BTNL3 makes physical contact with V4γ-bearing TCRs on γδ T cells.
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Receptores de Antígenos de Linfocitos T gamma-delta , Linfocitos T , Butirofilinas , Membrana Celular , Humanos , LigandosRESUMEN
Human farnesyl pyrophosphate synthase (Homo sapiens FPPS, HsFPPS) is a target for treating bone resorption diseases and some cancers. HsFPPS is potently inhibited by bisphosphonates, but due to poor cell penetration and distribution in soft tissue, there is currently interest in the development of non-bisphosphonate inhibitors as cancer therapeutics. Here, we report the discovery and development of HsFPPS inhibitors based on the phenolic diterpene carnosic acid (CA), an antimicrobial found in rosemary and sage, which showed better cellular anticancer activities than the bisphosphonate drug zoledronate in pancreatic cancer cell lines, as well as an HsFPPS-dependent mechanism of action. Hit-to-lead optimization of CA improved HsFPPS inhibition by >100-fold. A slow dissociation inhibition pattern and a noncompetitive allosteric binding mode were found, and cellular mechanism-of-action studies showed that these inhibitors inhibit tumor cell growth primarily by inhibiting HsFPPS, leading to downregulation of Ras prenylation and cell apoptosis. The discovery of this series of compounds together with proof-of-mechanism in pancreatic cancer cells may pave the way for targeting HsFPPS in soft tissue cancers using natural-product-derived inhibitors.