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Cerebral ischemia reperfusion (I/R) is a therapeutic strategy for ischemia; however, it usually causes injury by the aspect of inflammation and neuron apoptosis. This investigation aims to investigate the protective effects of phytic acid (IP6) for cerebral I/R injury in vitro. PC-12 cells under Oxygen and glucose deprivation/reperfusion (OGD/R) were performed to mimic cerebral I/R. IP6 was pretreated before PC-12 cells under OGD/R treatment. The data showed that IP6 activated the expression of sestrin2 in OGD/R injured PC-12 cells. IP6 inhibited OGD/R induced inflammation, oxidative stress, and apoptosis by activating sestrin2. Besides, p38 MAPK may mediate the effects of sestrin2 activated by IP6. Therefore, IP6 can be a potential drug to prevent neurological damage in cerebral I/R injury.
Assuntos
Antioxidantes/metabolismo , Isquemia Encefálica/metabolismo , Fármacos Neuroprotetores/farmacologia , Proteínas Nucleares/metabolismo , Ácido Fítico/farmacologia , Traumatismo por Reperfusão/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Glucose/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Proteínas Nucleares/genética , Estresse Oxidativo/efeitos dos fármacos , Oxigênio/metabolismo , Células PC12 , Ratos , Transdução de Sinais/genética , Transfecção , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Current -omics methods allow the collection of a large amount of information that helps in describing the microbial diversity in nature. Here, and as a result of a culturomic approach that rendered the collection of thousands of isolates from 5 different hypersaline sites (in Spain, USA and New Zealand), we obtained 21 strains that represent two new Salinibacter species. For these species we propose the names Salinibacter pepae sp. nov. and Salinibacter grassmerensis sp. nov. (showing average nucleotide identity (ANI) values < 95.09% and 87.08% with Sal. ruber M31T, respectively). Metabolomics revealed species-specific discriminative profiles. Sal. ruber strains were distinguished by a higher percentage of polyunsaturated fatty acids and specific N-functionalized fatty acids; and Sal. altiplanensis was distinguished by an increased number of glycosylated molecules. Based on sequence characteristics and inferred phenotype of metagenome-assembled genomes (MAGs), we describe two new members of the genus Salinibacter. These species dominated in different sites and always coexisted with Sal. ruber and Sal. pepae. Based on the MAGs from three Argentinian lakes in the Pampa region of Argentina and the MAG of the Romanian lake Fara Fund, we describe the species Salinibacter pampae sp. nov. and Salinibacter abyssi sp. nov. respectively (showing ANI values 90.94% and 91.48% with Sal. ruber M31T, respectively). Sal. grassmerensis sp. nov. name was formed according to the rules of the International Code for Nomenclature of Prokaryotes (ICNP), and Sal. pepae, Sal. pampae sp. nov. and Sal. abyssi sp. nov. are proposed following the rules of the newly published Code of Nomenclature of Prokaryotes Described from Sequence Data (SeqCode). This work constitutes an example on how classification under ICNP and SeqCode can coexist, and how the official naming a cultivated organism for which the deposit in public repositories is difficult finds an intermediate solution.
Assuntos
Bacteroidetes , Ácidos Graxos , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Filogenia , Ácidos Graxos/análise , DNA Bacteriano/genética , Técnicas de Tipagem BacterianaRESUMO
OBJECTIVE: To evaluate the feasibility of non-sticky thermosensitive liquid embolic material chitosan/ß-glycerophosphate sodium (C/GP) and fibroblast embolization in rete mirabile (REM) for preparing the model of cerebral arteriovenous malformation (cAVM); to study the method of microcatheter injection of C/GP-gel system; and to observe the embolization effect and histological changes of REM. METHODS: A total of 26 domestic pigs were grouped and prepared designed models, followed by different treatment methods using C/GP. C/GP embolization of the REM were performed. The brain samples were obtained after week 6's angiography and finally, subjected to H&E staining for histological examination. RESULTS: In 26 pig models, 25 pigs were successfully modeled, and 1 pig had convulsions and died during the modeling process. After embolization, angiography showed that the embolized REM was no longer developed while there was no adhesion between the tip of the microcatheter and the embolization agent. No recanalization was found in week 2 and week 6's angiography. Histological examination: the hydrogel was uniformly dispersed in REM, and REM was completely embolized. The texture was hard. REM was filled by gel and fibroblasts, the intima of the wall was clearly visible, and the smooth muscle layer was intact. No exfoliation and necrosis of the vessel wall were observed, and no inflammatory reaction was observed around the blood vessel. CONCLUSIONS: Our study provided sufficient evidence to suggest that C/GP may be a novel liquid embolic material for the endovascular treatment of cAVM. C/GP and fibroblasts can be used in the embolization of cAVM and may have broad application as an ideal embolization material for the treatment of cAVM.
Assuntos
Quitosana , Embolização Terapêutica , Malformações Arteriovenosas Intracranianas , Suínos , Animais , Embolização Terapêutica/métodos , Malformações Arteriovenosas Intracranianas/terapia , FibroblastosRESUMO
Bacteria in root nodules of legumes play important roles in promoting plant growth. In this study, we investigated root nodule-associated bacteria isolated from leguminous plants along an elevation gradient on the northern slope of the Kunlun Mountains, China, using a cultivation approach. In total, 300 isolates were obtained from seven legume species within six ecological zones. Isolates were identified based on 16S rRNA gene phylogenetic analysis and potential rhizobia were further identified using a recA gene phylogeny. Among the isolates, Bacillales (particularly Bacillus) were the dominant isolates from all host legumes and all elevations (63.5%), followed by Rhizobiales (13%) and Pseudomonadales (11.7%). Less than 3% of the isolates belonged to Burkholderiales, Paenibacillales, Enterobacteriales, Actinomycetales, Sphingomonadales, Xanthomonadales, Chitinophagales, Brevibacillales, Staphylococcales, or Mycobacteriales. A few elevation-specific patterns emerged within the Bacillales and Pseudomonadales. For example, isolates related to the psychrotroph Bacillus psychrosaccharolyticus were only isolated from the highest elevation sites (>3,500 m) whereas those related to the mesophile Bacillus endophyticus were only isolated from lowest elevation sites (1,350 m), suggestive of a role of soil temperature in their distribution. Similarly, isolates related to Pseudomonas brassicacearum were the dominant Pseudomonadales isolates, but they were only isolated from middle and low elevations (<3,200 m). A total of 39 isolates belonged to the Rhizobiales, 36 of which were confirmed to the genus level using the recA gene. In all, Rhizobiales isolates were obtained from five different host legumes spanning the entire elevation gradient. Those from the low-elevation Qira Desert-Oasis Transition Zone (1,350-1,960 m) suggested some patterns of host preference. For example, most isolates from Albizia julibrissin formed a monophyletic group related to Rhizobium lemnae and most from Alhagi sparsifolia were closely related to Ensifer kummerowiae. In general, this study shows that most bacteria associated with root nodules of legumes are widely distributed in distinct ecological zones within a single geographic region but suggests that both climate and host interactions may influence their distributions.
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PSMA3 antisense RNA 1 (PSMA3AS1), a long noncoding RNA, promotes the progression of esophageal squamous cell carcinoma. However, no study to date has explored the expression or roles of PSMA3AS1 in nonsmall cell lung carcinoma (NSCLC). The present study examined the expression profile and role of PSMA3AS1 in NSCLC. It also aimed to identify how PSMA3AS1 promotes the malignant phenotype of NSCLC cells. PSMA3AS1 expression in NSCLC tissues and cell lines was measured by reverse transcriptionquantitative polymerase chain reaction. Cell Counting Kit8, cell apoptosis, Transwell migration and invasion, and xenograft tumor assays were conducted to study the effects of PSMA3AS1 on the aggressive phenotype of NSCLC cells. Furthermore, bioinformatics analysis, RNA immunoprecipitation, luciferase reporter assay, western blotting, and rescue experiments were used to elucidate the interaction among PSMA3AS1, microRNA4093p (miR4093p), and spindlin 1 (SPIN1) in NSCLC cells. In the present study, high levels of PSMA3AS1 were confirmed in both NSCLC tissues and cell lines. An increased PSMA3AS1 level was correlated with advanced tumornodemetastasis stage and increased lymph node metastasis. Patients with NSCLC with high PSMA3AS1 levels had shorter overall survival than those with low PSMA3AS1 levels. PSMA3AS1 depletion significantly decreased NSCLC cell proliferation, migration, and invasion, as well as substantially increased cell apoptosis in vitro. Furthermore, PSMA3AS1 deficiency decreased NSCLC tumor growth in vivo. Through molecular mechanism assays, it was revealed that PSMA3AS1 acted as a molecular sponge for miR4093p and consequently increased SPIN1 expression. Notably, rescue experiments revealed that the inhibition of miR4093p or restoration of SPIN1 expression abrogated the effects of PSMA3AS1 knockdown in NSCLC cells. Collectively, PSMA3AS1 functioned as an oncogenic long noncoding RNA in NSCLC. PSMA3AS1 sponged miR4093p and thus increased SPIN1 expression, promoting the aggressive phenotype of NSCLC cells.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Proteínas de Ciclo Celular/genética , MicroRNAs/genética , Proteínas Associadas aos Microtúbulos/genética , Fosfoproteínas/genética , RNA Longo não Codificante/genética , Células A549 , Animais , Apoptose/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica , Xenoenxertos , Humanos , Metástase Linfática/genética , Metástase Linfática/patologia , Masculino , Camundongos , Pessoa de Meia-Idade , Complexo de Endopeptidases do Proteassoma/genéticaRESUMO
The present study was designed to investigate the function of matrix metalloproteinase9 (MMP9) in human glioma cells and the potential regulatory mechanisms. Reverse transcriptionquantitative polymerase chain reaction was used to analyze the expression of MMP9 and microRNA34a (miR34a) in the plasma of patients with glioma and healthy volunteers. MTT and Transwell assays were used to assess cell growth and migration, respectively. AnnexinV/propidium iodide staining was used to measure cell apoptosis. In addition, MMP9 expression was measured using western blot analysis. In patients with glioma, MMP9 expression was increased, while miR34a expression was suppressed, compared with the normal group. Overall survival (OS) and diseasefree survival (DFS) of patients with high MMP9 expression were decreased compared with those with low MMP9 expression. OS and DFS of patients with low miR34a expression were decreased compared with those with high miR34a expression. Downregulation of miR34a promoted cell growth and migration, and inhibited apoptosis in U251MG glioma cells. However, overexpression of miR34a inhibited cell growth and migration, and induced apoptosis in glioma cells. Furthermore, downregulation of miR34a using antimiR34a induced MMP9 protein expression in glioma cells; whereas, overexpression of miR34a suppressed MMP9 protein expression in glioma cells. SB3CT, an inhibitor of MMP9, attenuated the effects of miR34a mimic on glioma cells. Together, these results indicated that miR34a inhibited cell growth and migration in human glioma cells by regulating MMP9.
Assuntos
Proliferação de Células/genética , Glioma/genética , Metaloproteinase 9 da Matriz/genética , MicroRNAs/genética , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Regulação Neoplásica da Expressão Gênica/genética , Glioma/patologia , HumanosRESUMO
The aim of the present study was to assess the expression status of matrix metalloproteinase (MMP)-28 and to investigate its molecular mechanisms in glioma cells. MicroRNA (miRNA) reverse transcription-quantitative polymerase chain reaction was used to analyze the expression of MMP-28 and transforming growth factor (TGF)-ß expression in glioma patients and healthy volunteers. MTT and Transwell assays were conducted to determine cell growth and metastasis, respectively. Annexin V/propidium iodide staining was also employed to measure cell apoptosis. MMP-28 and TGF-ß protein expression were measured using western Blot analysis. The results indicated that MMP-28 and TGF-ß expression was downregulated in glioma patients, when compared with the normal group. Overall survival and disease-free survival of patients with a low expression of MMP-28 were lower than those with high MMP-28 expression. Overexpression of MMP-28 induced TGF-ß protein expression, while downregulation of MMP-28 suppressed TGF-ß protein expression in glioma cell. The downregulation of MMP-28 reduced the cell growth and apoptosis of glioma cell via the suppression of TGF-ß. By contrast, upregulation of MMP-28 induced cell growth and reduced the apoptosis of glioma cells by activating TGF-ß. In addition, the TGF-ß inhibitor attenuated the effects of MMP-28 in glioma cells. Collectively, the results indicated that MMP-28 was able to induce TGF-ß in human glioma cells.
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The aim of the present study was to evaluate the feasibility of the non-adhesive temperature-sensitive liquid embolic material, chitosan/ß-glycerophosphate (C/GP), in embolizing the basicranial rete mirabile (REM) in a swine model of cerebral arteriovenous malformation (cAVM). A total of 24 domestic swines were used as the experimental animals, among which 12 pigs underwent direct embolization of one side of the REM, while the other 12 pigs underwent embolization of the bilateral REM following anastomosis of the carotid artery and jugular vein. A super-selective microcatheter was introduced into the REM during the embolization procedure, and the C/GP hydrogel was injected until an image of the REM disappeared in the angiography examination. Further angiography examinations were performed after 2 and 6 weeks, and histological examination of the REM was performed after 6 weeks. Of the 24 domestic swines, 23 cases underwent successful thrombosis. Convulsions occurred in one case and that pig died during the embolization procedure. Following embolization, the angiography observations revealed that the embolized REM was no longer able to be developed, and adhesion of the microcatheter tip with the embolic agent did not occur. In addition, no apparent revascularization was observed in the angiography examinations performed at weeks 2 and 6. Therefore, the current preliminary study indicated that use of the non-adhesive temperature-sensitive embolic material was feasible for the embolization of cAVM; thus, C/GP may be used as an ideal embolic material for the treatment of cAVM.
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This study aimed to investigate the inhibitory roles of a polysaccharide extract from Inonotus obliquus on U251 human neurogliocytoma cells cultured in vitro. After administering the polysaccharide extract from I. obliquus to U251 cells cultivated in vitro, methyl thiazolyl tetrazoliym assay was performed to measure the inhibitory effects of the extract on tumor cell proliferation. The expression of the apoptosis-related proteins Bcl-2 and caspase-3 were determined by Western blotting. Different concentrations of I. obliquus extract (25, 50, 100, 200, and 500 µg/mL) were added to U251 cells at 24, 48, and 72 hours. Methyl thiazolyl tetrazoliym assay showed that the inhibition ratio increased with increased extract concentration and prolonged treatment duration. The I. obliquus extract sharply decreased the expression of Bcl-2 but dramatically increased the expression of caspase-3. This function was gradually enhanced with increased drug concentration and prolonged treatment duration. The I. obliquus extract can inhibit the proliferation of tumor cells. This inhibition function is closely related to the downregulation of Bcl-2 and the upregulation of caspase-3.