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Two related anaerobic strains, designated as SWB101512T and SWB19611, were isolated from the bronchoalveolar lavage fluid of two lung cancer patients. Cells were Gram-stain-positive, non-motile and non-spore-forming. Growth could be observed at 26-45 °C (optimum, 37 °C), pH 5.0-8.5 (optimum, pH 7.0) and with 0.5-2.0â% (v/w) NaCl (optimum, 1.0%). The 16S rRNA gene sequences of SWB101512T and SWB19611 showed the highest similarities to Denitrobacterium detoxificans DSM 21843T (91.1 and 91.3â%, respectively). The phylogenetic tree based on the 16S rRNA gene sequences and the core genome sequences demonstrated that the two strains clustered together and formed a distinct lineage within the family Eggerthellaceae. The DNA G+C contents of strains SWB101512T and SWB19611 were 62.0 and 61.9âmol%, respectively. The predominant cellular fatty acids of strains SWB101512T and SWB19611 were C16â:â0 DMA (27.8 and 28.8â%, respectively). The respiratory menaquinone in both strains was menaquinone 6 and the polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, two phospholipids, three glycolipids and three unidentified lipids. Based on evidence from phenotypic, chemotaxonomic and genomic analyses, a new genus and species belonging to the family Eggerthellaceae, named Curtanaerobium respiraculi gen. nov., sp. nov. is proposed. The type strain is SWB101512T (=GDMCC 1.2991T=JCM 35330T).
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Actinobacteria , Ácidos Graxos , Humanos , Ácidos Graxos/química , Filogenia , Composição de Bases , RNA Ribossômico 16S/genética , Anaerobiose , Líquido da Lavagem Broncoalveolar , DNA Bacteriano/genética , Análise de Sequência de DNA , Técnicas de Tipagem Bacteriana , Fosfolipídeos/química , Bactérias Anaeróbias/genética , Actinobacteria/genética , ChinaRESUMO
OBJECTIVE: Due to the rarity and non-specificity of symptoms, gastric metastases are often misdiagnosed, and patients are not treated promptly. The aim of this study was to study the clinicopathological features and differential diagnosis of gastric metastases. METHODS: From 2004 to 2021, 14 patients were diagnosed with gastric metastases not resulting from direct invasion (GMNDI) in our hospital, and their imaging and clinicopathological features were analyzed. RESULTS: PET-CT examination showed hypermetabolic nodules in the stomach. Under gastroscopy, GMNDI showed eminence, nodular or vegetable pattern mass, and ulcer. Microscopically, GMNDI showed similar pathological features and immunophenotypes to the primary tumor. In our study, the most common primary tumors were malignant melanoma (4 cases), small cell lung cancer (3 cases), and hepatocellular carcinoma (3 cases). Immunohistochemistry contributed to the pathological diagnosis and differential diagnosis of gastric metastases. Malignant melanoma expressed HMB45, MelanA, and S-100; small cell lung cancer expressed TTF-1, CD56, and CgA; hepatocellular carcinoma expressed GPC-3, hepatocyte, and Sall4. In a few cases, tumor cells may lose immune markers during metastasis. Therefore, it is necessary to combine medical history, imaging examination, and other clinical diagnosis methods in the pathological diagnosis. CONCLUSION: An in-depth understanding of GMNDI is conducive to better diagnosis and treatment planning for gastric metastases and subsequent improvement in patient prognosis.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Neoplasias Pulmonares , Melanoma , Carcinoma de Pequenas Células do Pulmão , Humanos , Diagnóstico Diferencial , Carcinoma Hepatocelular/diagnóstico , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Estômago , Melanoma/diagnóstico , Neoplasias Hepáticas/diagnóstico , Neoplasias Pulmonares/diagnóstico , Melanoma Maligno CutâneoRESUMO
Metal organic frameworks (MOFs)-derived porous carbon is proposed as a promising candidate to develop novel, tailorable structures as polysulfides immobilizers for lithium-sulfur batteries because of their high-efficiency electron conductive networks, open ion channels, and abundant central ions that can store a large amount of sulfur and trap the easily soluble polysulfides. However, most central ions in MOFs-derived carbon framework are encapsulated in the carbon matrix so that their exposures as active sites to adsorb polysulfides are limited. To resolve this issue, highly dispersed TiO2 nanoparticles are anchored into the cobalt-containing carbon polyhedras that are converted from ZIF-67. Such a type of TiO2 and Co nanoparticles-decorated carbon polyhedras (CCo/TiO2 ) provide more exposed active sites and much stronger chemical trapping for polysulfides, hence improving the sulfur utilization and enhancing reaction kinetics of sulfur-containing cathode simultaneously. The sulfur-containing carbon polyhedras decorated with TiO2 nanoparticles (S@CCo/TiO2 ) show a significantly improved cycling stability and rate capability, and deliver a discharge capacity of 32.9% higher than that of TiO2 -free S@CCo cathode at 837.5 mA g-1 after 200 cycles.
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OBJECTIVE: To study differential expression of microRNA (miRNA) in peripheral blood mononuclear cells (PBMNC) from patients with different types of aplastic anemia (AA) and explore the role of miRNA in the pathogenesis of AA. METHODS: miRNA microarray were used to determine the differential expression profile of miRNA in PBMNC from patients with AA. Real-time quantitative polymerase china reaction (RQ-PCR) was used to verify the differential expression of miRNA. Candidate miRNA were analyzed with bioinformatics tools. RESULTS: Compared with the normal controls, 6 miRNAs were up-regulated and 10 were down-regulated in patients with severe aplastic anemia (SAA), while 24 miRNAs were up-regulated and 12 were down-regulated in patients with chronic non-severe aplastic anemia (CAA). Compared with CAA patients, 4 miRNAs were up-regulated and 11 were down-regulated in SAA patients. Compared with normal controls, 3 miRNAs were up-regulated and 4 were down-regulated in both SAA and CAA patients. As verified by RQ-PCR, expression of miR-155-5p and miR-1260b were increased in both CAA and SAA patients compared with the normal controls (P<0.01). The expression of miR-155-5p and miR-1260b of CAA patients were higher than that of SAA patients (P<0.01). Bioinformatics analysis showed that target genes of miR-155-5p and miR-1260b may be involved in regulation of cell metabolism, gene expression and transcription, TNF signaling pathway, B cell receptor signaling pathway, Fc gamma R-mediated phagocytosis and other signaling process. CONCLUSION: There are characteristic differential expression profiles of miRNA in PBMNC from CAA and SAA patients, in which miRNA-155-5p and miRNA-1260b are both up-regulated. The common target gene predicted for miRNA-155-5p and miRNA-1260b is ETS1. miRNA-155-5p and miRNA-1260b may act synergistically to inhibit the expression of ETS1 and promote differentiation of Th17, therefore play an important role in the pathogenesis of AA.
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Anemia Aplástica/genética , Leucócitos Mononucleares/metabolismo , MicroRNAs/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Anemia Aplástica/metabolismo , Anemia Aplástica/fisiopatologia , Diferenciação Celular , China , Biologia Computacional , Feminino , Perfilação da Expressão Gênica , Humanos , Masculino , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Células Th17/citologia , Células Th17/metabolismo , Adulto JovemRESUMO
In this research, Ru-doped TiO2 nanotube arrays (Ru-TNTA) were prepared by anodizing TiRu alloys, and the effects of annealing temperature, Ru content and test temperature on their performances for the hydrogen evolution reaction (HER) and the oxygen evolution reaction (OER) were investigated. The results show that the unannealed Ru-TNTA (a-Ru-TNTA) exhibits superior activity for the HER, and the Ru-TNTA annealed at 450 °C (c-Ru-TNTA) shows excellent activity for the OER. The Ru content of TiRu impacts the electrochemically active surface area (ECSA) and the charge transfer resistance (Rct) significantly. When the Ru content of Ru-TNTA is 6%, its performance is optimal. Moreover, the electrocatalytic activity of Ru-TNTA improves with increasing test temperature, and the overpotentials of a-Ru-TNTA and c-Ru-TNTA at 80 °C are 19 mV and 227 mV (10 mA cm-2), respectively. Ru-TNTA exhibits excellent electrocatalytic performance for water splitting and good stability, which provides a new idea for the preparation of advanced bifunctional electrocatalysts for water splitting.
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The special blood circulation, anatomy, and tissue structure of the spine may lead to significant differences in pathological features and drug resistance between spinal tuberculosis and pulmonary tuberculosis. Here, we collected 168 spinal tuberculosis cases and 207 pulmonary tuberculosis cases, and compared their clinical and pathological features as well as drug resistance. From the anatomical location, the highest incidence was of lumbar tuberculosis, followed by thoracic tuberculosis. PET-CT scans showed increased FDG uptake in the diseased vertebrae, discernible peripheral soft tissue shadow, visible internal capsular shadow, and an abnormal increase in FDG uptake. MRI showed infectious lesions in the diseased vertebral body, formation of paravertebral and bilateral psoas muscle abscess, and edema of surrounding soft tissues. As with control tuberculosis, the typical pathological features of spinal tuberculosis were chronic granulomatous inflammation with caseous necrosis. The incidence of granulomas was not statistically different between the groups. However, the proportions of caseous necrosis, acute inflammation, abscess, exudation, and granulation tissue formation in the spinal tuberculosis group were all significantly increased relative to the control tuberculosis group. Compared to the control tuberculosis group, the incidences of resistance to rifampicin (RFP) + isoniazid (INH) + streptomycin (STR) and INH + ethambutol (EMB) were lower in the spinal tuberculosis group, while the incidences of resistance to RFP + INH + EMB and RFP + EMB were higher. Moreover, we also found some differences in drug-resistance gene mutations. In conclusion, there are noticeable differences between spinal Mycobacterium tuberculosis and pulmonary tuberculosis in pathological characteristics, drug resistance, and drug resistance gene mutations.
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Tumor Metabolism is strongly correlated with prognosis. Nevertheless, the prognostic and therapeutic value of metabolic-associated genes in BCa patients has not been fully elucidated. First, in this study, metabolism-related differential expressed genes DEGs with prognostic value in BCa were determined. Through the consensus clustering algorithm, we identified two molecular clusters with significantly different clinicopathological features and survival prognosis. Next, a novel metabolism-related prognostic model was established. Its reliable predictive performance in BCa was verified by multiple external datasets. Multivariate Cox analysis exhibited that risk score were independent prognostic factors. Interestingly, GSEA enrichment analysis of GO, KEGG, and Hallmark gene sets showed that the biological processes and pathways associated with ECM and collagen binding in the high-risk group were significantly enriched. Notely, the model was also significantly correlated with drug sensitivity, immune cell infiltration, and immunotherapy efficacy prediction by the wilcox rank test and chi-square test. Based on the 7 immune infiltration algorithm, we found that Neutrophils, Myeloid dendritic cells, M2 macrophages, Cancer-associated fibroblasts, etc., were more concentrated in the high-risk group. Additionally, in the IMvigor210, GSE111636, GSE176307, or our Truce01 (registration number NCT04730219) cohorts, the expression levels of multiple model genes were significantly correlated with objective responses to anti-PD-1/anti-PD-L1 immunotherapy. Finally, the expression of interested model genes were verified in 10 pairs of BCa tissues and para-carcinoma tissues by the HPA and real-time fluorescent quantitative PCR. Altogether, the signature established and validated by us has high predictive power for the prognosis, immunotherapy responsiveness, and chemotherapy sensitivity of BCa.
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Neoplasias da Bexiga Urinária , Humanos , Neoplasias da Bexiga Urinária/genética , Prognóstico , Algoritmos , Análise por Conglomerados , Consenso , Microambiente TumoralRESUMO
BACKGROUND: Colorectal cancer (CRC) is the third most common cancer in the world, and a strong relationship exists between CRC and gut microbiota, which affects the occurrence, development, and metastasis of cancer. Bioinformatics-based analyses revealed that the abundance of Parvimonas micra (P. micra) in the feces of patients with cancer is significantly higher than that in healthy people. Therefore, an important relationship may exist between P. micra and CRC. METHODS: We first confirmed that P. micra can promote the proliferation of cell lines through cell experiments and mouse models. Then we selected the signaling pathways and content of exosomes to promote the development of CRC by transcriptomics and microRNA sequencing. Finally, we confirmed that P. micra promoted CRC development through miR-218-5p/Ras/ERK/c-Fos pathway through the in vivo and in vitro experiments. RESULTS: First, it was confirmed by in vitro and in vivo experiments that P. micra can promote the development of CRC. Transcriptome analysis after the coincubation of bacteria and cells revealed that P. micra promoted cell proliferation by activating the Ras/ERK/c-Fos pathway. Furthermore, microRNA sequencing analysis of the cells and exosomes showed that miR-218-5p and protein tyrosine phosphatase receptor R (PTPRR) were the key factors involved in activating the Ras/ERK/c-Fos pathway, and the miR-218-5p inhibitor was used to confirm the role of microRNA in xenograft mice. CONCLUSION: This experiment confirmed that P. micra promoted the development of CRC by upregulating miR-218-5p expression in cells and exosomes, inhibiting PTPRR expression, and ultimately activating the Ras/ERK/c-Fos signaling pathway.
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Neoplasias Colorretais , Firmicutes , MicroRNAs , Animais , Humanos , Camundongos , Linhagem Celular Tumoral , Proliferação de Células/genética , Neoplasias Colorretais/microbiologia , Neoplasias Colorretais/patologia , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , MicroRNAs/metabolismo , Transdução de Sinais , Firmicutes/patogenicidadeRESUMO
Recently, microbiota dysbiosis in lung cancer has attracted immense attention. Studies on lung microbes are mostly based on sequencing, which has left the potentially functional bacteria with extremely low abundance uncovered. In this study, we characterized and compared the lung and oral cavity microbiotas using culturomics and 16S rRNA gene sequencing. Of the 198 bacteria identified at the species level from bronchoalveolar lavage fluid (BALF) samples, Firmicutes was predominant (39.90%). Twenty bacterial species isolated from BALF samples were present in at least half of the patients and were also highly abundant in oral samples. Of all isolated strains, Streptococcus and Veillonella were highly dominant. The abundance of Prevotella and Veillonella decreased from the oral cavity to the lung, whereas that of Pseudomonas increased. Linear discriminant analysis effect size demonstrated that Prevotella was more abundant in the healthy samples than in the cancerous ones, which is in accordance with the isolation of Prevotella oralis only from the healthy group using culturomics. Moreover, Gemella sanguinis and Streptococcus intermedius were isolated only from the non-small-cell lung cancer (NSCLC) group, and 16S rRNA gene sequencing showed that they were higher in the NSCLC than in the small-cell lung cancer group. Furthermore, while Bacillus and Castellaniella were enriched in lung adenocarcinoma, Brucella was enriched in lung squamous cell carcinoma. Overall, alterations were observed in the microbial community of patients with lung cancer, whose diversity might be site and pathology dependent. Using culturomics and 16S rRNA gene amplicon sequencing, this study has provided insights into pulmonary and oral microbiota alterations in patients with lung cancer. IMPORTANCE The relationship between lung microbiota and cancer has been explored based on DNA sequencing; however, culture-dependent approaches are indispensable for further studies on the lung microbiota. In this study, we applied a comprehensive approach combining culturomics and 16S rRNA gene amplicon sequencing to detect members of the microbiotas in saliva and BALF samples from patients with unilateral lobar masses. We found alterations in the microbial community of patients with lung cancer, whose diversity might be site and pathology dependent. These features may be potential bacterial biomarkers and new targets for lung cancer diagnosis and treatment. In addition, a lung and oral microbial biobank from lung cancer patients was established, which represents a useful resource for studies of host-microbe interactions.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Microbiota , Humanos , RNA Ribossômico 16S/genética , Genes de RNAr , Pulmão/microbiologia , Microbiota/genética , BactériasRESUMO
In recent years, the incidence of tuberculosis (TB) and mortality caused by the disease have been decreasing. However, the number of drug-resistant tuberculosis patients is increasing rapidly year by year. Here, a total of 380 Mycobacterium tuberculosis (MTB)-positive formalin-fixed and paraffin-embedded tissue (FFPE) specimens diagnosed in the Department of Pathology of the Eighth Medical Center, Chinese PLA General Hospital were collected. Among 380 cases of MTB, 85 (22.37%) were susceptible to four anti-TB drugs and the remaining 295 (77.63%) were resistant to one or more drugs. The rate of MDR-TB was higher in previously treated cases (52.53%) than in new cases [(36.65%), p < 0.05]. Of previously treated cases, the rate of drug resistance was higher in females than in males (p < 0.05). Among specimens obtained from males, the rate of drug resistance was higher in new cases than in previously treated cases (p < 0.05). Of mutation in drug resistance-related genes, the majority (53/380, 13.95%) of rpoB gene carried the D516V mutation, and 13.42% (51/380) featured mutations in both the katG and inhA genes. Among the total specimens, 18.68% (71/380) carried the 88 M mutation in the rpsL gene, and the embB gene focused on the 306 M2 mutation with a mutation rate of 19.74%. Among the resistant INH, the mutation rate of −15 M was higher in resistance to more than one drug than in monodrug-resistant (p < 0.05). In conclusion, the drug resistance of MTB is still very severe and the timely detection of drug resistance is conducive to the precise treatment of TB.
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BACKGROUND: As a FAD (Flavin Adenine Dinucleotide) - dependent histone demethylase discovered in 2004, LSD1 (lysine-specific demethylase 1) was reported to be overexpressed in diverse tumors, regulating target genes transcription associated with cancer development. Hence, LSD1 targeted inhibitors may represent a new insight in anticancer drug discovery. For these reasons, researchers in both the pharmaceutical industry and academia have been actively pursuing LSD1 inhibitors in the quest for new anti-cancer drugs. OBJECTIVES: This review summaries patents about LSD1 inhibitors in recent 5 years in the hope of providing a reference for LSD1 researchers to develop new modulators of LSD1 with higher potency and fewer adverse effects. METHODS: This review collects LSD1 inhibitors disclosed in patents since 2016. The primary ways of patent searching are Espacenet®, Google Patents, and CNKI. RESULTS: This review covers dozens of patents related to LSD1 inhibitors in recent five years. The compound structures are mainly divided into TCP (Tranylcypromine) derivatives, imidazole derivatives, pyrimidine derivatives, and other natural products and peptides. Meanwhile, the compounds that have entered the clinical phase are also described. CONCLUSION: Most of the compounds in these patents have been subjected to activity analysis with LSD1 and multi-cell lines, showing good antitumor activity in vitro and in vivo. These patents exhibited the structural diversity of LSD1 inhibitors and the potential of natural products as novel LSD1 inhibitors.
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Antineoplásicos , Lisina , Antineoplásicos/química , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/uso terapêutico , Histona Desmetilases/química , Humanos , Patentes como Assunto , Tranilcipromina/química , Tranilcipromina/farmacologiaRESUMO
Dual target compounds have become a hot spot in the treatment of cancer in recent years. Histone lysine specific demethylase 1 (LSD1) is identified as histone demethylase and acts as a key regulator involved in many other cellular activities through its demethylation function. We have reported a triazolo [1,5-α] pyrimidine-based DCN1(defective in cullin neddylation protein 1) inhibitor compound 383 (IC50 = 11 nM) which could selectively inhibit Cullin 3/1 neddylation in MGC-803 cells. In this research, we investigated that compound 383 could target LSD1 and inhibit the biological function of LSD1 in MGC-803 cells (IC50 = 0.53 µM). We found that compound 383 could induce the degradation of LSD1 and inhibit MGC-803 cell proliferation, migration and invasion in a dose-dependent manner. Compound 383 could cause cell cycle arrest at G2/M phase by down-regulating the expression of LSD1. In addition, compound 383 could significantly reverse epithelial-mesenchymal transition (EMT) through increase H3K4me methylation at E-cadherin promotor. Furthermore, the in vivo inhibitory effect of compound 383 without obvious toxicity was confirmed in nude mouse transplanted MGC-803 tumor cells model. Collectively, these results suggest that the DCN1 inhibitor compound 383 exhibits antiproliferative activity in gastric cancer cells by targeting LSD1 which promotes compound 383 as a good starting point for the development of dual-target therapeutics for gastric cancer.
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Inibidores Enzimáticos , Histona Desmetilases , Peptídeos e Proteínas de Sinalização Intracelular , Neoplasias Gástricas , Linhagem Celular Tumoral , Descoberta de Drogas , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/uso terapêutico , Histona Desmetilases/antagonistas & inibidores , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/antagonistas & inibidores , Neoplasias Gástricas/tratamento farmacológico , Relação Estrutura-AtividadeRESUMO
The recurrence rate of lymphoma is very high, and tumor stem cells may be an important mechanism. Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) can inhibit antitumor immunity and promote cancer progression, but its role and mechanism in lymphoma are still unclear. Here we collected lymphoma tissue and peripheral blood from patients with diffuse large B-cell lymphoma (DLBCL). Results showed that CTLA-4 expression and CD44+ cell in the high-risk group were significantly higher than that in the low-risk group. Correlation analysis showed that CTLA-4 expression positively correlated with CD44+ cell in lymphoma tissue and regulatory T (Treg) cells in lymphocytes. In vitro experiment showed that CTLA-4 increased the ratio of lymphoma stem cells, and proliferation and invasion of lymphoma cells through TGF-ß pathway. Moreover, CTLA-4 enhanced the proliferation of Treg cells induced by lymphoma cells. Animal experiments showed that CTLA-4 can promote transplanted lymphoma growth. Immunohistochemistry results showed that both Ki-67 and CD44+ cells increased significantly in the CTLA-4 group. TGF-ß neutralization can significantly block these effects of CTLA-4. In conclusion, CTLA-4 promoted DLBCL progression through lymphoma stem cell enrichment and immunosuppression.
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High cholesterol is one of the important factors inducing cardiovascular and cerebrovascular diseases. Drug therapy is the main method for reducing cholesterol, but has the disadvantages such as high cost and side effects. Studies have shown that intestinal bacteria play important roles in cholesterol metabolism. However, there are few reports on the screening and functional evaluation of cholesterol-lowering intestinal bacteria. In this study, 36 bile-tolerant bacteria were screened from healthy people stool through culturomics using bovine bile acid or artificial mixed bile acids as substrates. Taking Lactobacillus rhamnosus GG (LGG) as a positive control, three bile acid concentration groups (0 g/L, 0.3 g/L, 3 g/L) were set up to evaluate the cholesterol-lowering ability of bile-tolerant bacteria in vitro. Ten bacteria (including Proteus mirabilis, Providencia stuartii, Proteus vulgaris et al) were identified as the dominant cholesterol-lowering bacteria. Six of the above bacteria, Proteus mirabilis, Providencia stuartii, Proteus vulgaris, Proteus penneri, Wohlfahrtiimonas chitiniclastica, Providencia rettger, were evaluated for their ability to reduce triglycerides in vitro and tolerance to artificial gastric juice. Comparing with strain LGG, the six bacteria showed better triglyceride-lowering ability in vitro. With the decrease of pH value of artificial gastric juice and the increase of treatment time, the survival rate of six bacteria decreased. The above screening experiments and functional evaluation provide a basis for further development of potential cholesterol-lowering bacterial products.
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Proteus mirabilis , Providencia , Animais , Bovinos , Colesterol , Gammaproteobacteria , HumanosRESUMO
LSD1 and HDAC are physical and functional related to each other in various human cancers and simultaneous pharmacological inhibition of LSD1 and HDAC exerts synergistic anti-cancer effects. In this work, a series of novel LSD1/HDAC bifunctional inhibitors with a styrylpyridine skeleton were designed and synthesized based on our previously reported LSD1 inhibitors. The representative compounds 5d and 5m showed potent activity against LSD1 and HDAC at both molecular and cellular level and displayed high selectivity against MAO-A/B. Moreover, compounds 5d and 5m demonstrated potent antiproliferative activities against MGC-803 and HCT-116 cancer cell lines. Notably, compound 5m showed superior in vitro anticancer potency against a panel of gastric cancer cell lines than ORY-1001 and SP-2509 with IC50 values ranging from 0.23 to 1.56 µM. Compounds 5d and 5m significantly modulated the expression of Bcl-2, Bax, Vimentin, ZO-1 and E-cadherin, induced apoptosis, reduced colony formation and suppressed migration in MGC-803 cancer cells. In addition, preliminary absorption, distribution, metabolism, excretion (ADME) studies revealed that compounds 5d and 5m showed acceptable metabolic stability in human liver microsomes with minimal inhibition of cytochrome P450s (CYPs). Those results indicated that compound 5m could be a promising lead compound for further development as a therapeutic agent in gastric cancers via LSD1 and HDAC dual inhibition.
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Antineoplásicos/farmacologia , Desenho de Fármacos , Inibidores Enzimáticos/farmacologia , Histona Desacetilases/metabolismo , Histona Desmetilases/antagonistas & inibidores , Neoplasias Gástricas/tratamento farmacológico , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Histona Desmetilases/metabolismo , Humanos , Microssomos Hepáticos/química , Microssomos Hepáticos/metabolismo , Simulação de Acoplamento Molecular , Estrutura Molecular , Ratos , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
The two highly conserved NPA motifs (asparagine-proline-alanine, NPA) are the most important structural domains that play a crucial role in water-selective permeation in aquaporin water channels. However, the functions of NPA motifs in aquaporin (AQP) biogenesis remain largely unknown. Few AQP members with variations in NPA motifs such as AQP11 and AQP12 do not express in the plasma membrane, suggesting an important role of NPA motifs in AQP plasma membrane targeting. In this study, we examined the role of the two NPA motifs in AQP4 plasma membrane targeting by mutagenesis. We constructed a series of AQP4 mutants with NPA deletions or single amino acid substitutions in AQP4-M1 and AQP4-M23 isoforms and analyzed their expression patterns in transiently transfected FRT and COS-7 cells. Western blot analysis showed similar protein bands of all the AQP4 mutants and the wild-type AQP4. AQP4 immunofluorescence indicated that deletion of one or both NPA motifs resulted in defective plasma membrane targeting, with apparent retention in endoplasmic reticulum (ER). The A99T mutant mimicking AQP12 results in ER retention, whereas the A99C mutant mimicking AQP11 expresses normally in plasma membrane. Furthermore, the AQP4-M1 but not the M23 isoform with P98A substitution in the first NPA motif can target to the plasma membrane, indicating an interaction of N-terminal sequence of AQP4-M1 with the first NPA motif. These results suggest that NPA motifs play a key role in plasma membrane expression of AQP4 but are not involved in AQP4 protein synthesis and degradation. The NPA motifs may interact with other structural domains in the regulation of membrane trafficking during aquaporin biogenesis.
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Aquaporina 4/metabolismo , Oligopeptídeos/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Substituição de Aminoácidos , Animais , Aquaporina 4/genética , Células CHO , Células COS , Chlorocebus aethiops , Cricetinae , Cricetulus , Deleção de SequênciaRESUMO
Barium sulfate (BaSO4) is a material with high reflectance for preparing bifunctional sheets used in dry reagent chemical tests. In this study, bifunctional BaSO4 sheets with scattering power and high reflectance were prepared with BaSO4 microspheres sized 1.3~1.8 µm and cellulose acetate (CA). Factors such as the BaSO4 morphology, CA dosage, mixing time, surfactant, solid content and wet sheet thickness were investigated. Scanning electron microscopy (SEM), a dynamic contact angle test, light reflection detector and light reflection densitometer were employed to characterize the structure and properties of bifunctional BaSO4 sheets. The optimal conditions for preparing bifunctional BaSO4 sheets under natural drying conditions were as follows: mass ratio of CA to BaSO4 microspheres was 0.03:1; mixing acetone solution was used to form liquid stock puree; 35 µL Tween-80 was used to improve the hydrophilicity of bifunctional sheets; solid content was 54.9%; mixing 2 h then 450 µm thick sheets were cast on a glass plate using a film applicator. The light reflectance value of bifunctional BaSO4 sheets in the range of 400~800 nm was higher than 97%. Serum diffused in bifunctional BaSO4 sheets reacted in reagent sheets and formed uniform colorful spots. Considering the repeatability of spot proportion and light reflectance value, the sheet offered a uniform serum scattering power and good repeatability. Therefore, the bifunctional BaSO4 sheets are regarded as a promising material for dry chemical diagnostic reagents.
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Histone lysine specific demethylase 1 (LSD1) has become a potential therapeutic target for the treatment of cancer. Discovery and develop novel and potent LSD1 inhibitors is a challenge, although several of them have already entered into clinical trials. Herein, for the first time, we reported the discovery of a series of 5-cyano-6-phenylpyrimidine derivatives as LSD1 inhibitors using flavin adenine dinucleotide (FAD) similarity-based designing strategy, of which compound 14q was finally identified to repress LSD1 with IC50 = 183 nmol/L. Docking analysis suggested that compound 14q fitted well into the FAD-binding pocket. Further mechanism studies showed that compound 14q may inhibit LSD1 activity competitively by occupying the FAD binding sites of LSD1 and inhibit cell migration and invasion by reversing epithelial to mesenchymal transition (EMT). Overall, these findings showed that compound 14q is a suitable candidate for further development of novel FAD similarity-based LSD1 inhibitors.
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Lysine-specific demethylase 1 (LSD1), demethylase against mono- and di - methylated histone3 lysine 4, has emerged as a promising target in oncology. More specifically, it has been demonstrated as a key promoter in acute myeloid leukemia (AML), and several LSD1 inhibitors have already entered into clinical trials for the treatment of AML. In this paper, a series of new indole derivatives were designed and synthesized based on a lead compound obtained by a high-throughput screening with our in-house compound library. Among the synthetic compounds, 9e was characterized as a potent LSD1 inhibitor with an IC50 of 1.230⯵M and can inhibit the proliferation of THP-1â¯cells effectively. And most importantly, this is the first irreversible LSD1 inhibitor that is not derived from monoamine oxidase inhibitors. Hence, the discovery of 9e may serve as a proof of concept work for AML treatment.
Assuntos
Descoberta de Drogas , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/farmacologia , Histona Desmetilases/antagonistas & inibidores , Indóis/síntese química , Indóis/farmacologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Inibidores Enzimáticos/química , Inibidores Enzimáticos/metabolismo , Furanos/química , Ensaios de Triagem em Larga Escala , Histona Desmetilases/metabolismo , Humanos , Indóis/química , Indóis/metabolismo , Concentração Inibidora 50 , Leucemia Mieloide Aguda/patologia , Proteínas Recombinantes/efeitos dos fármacos , Proteínas Recombinantes/metabolismo , Relação Estrutura-AtividadeRESUMO
Polymorphonuclear neutrophil leukocyte (PMN) is an important member of the human immune cells. Recentyears, the recognition of the PMN function and the relationship between PMN and periodontitis have been updated. Besidesthe pathogens killing and phagocytosis, PMN also play an important role in immunoregulation and proresolving. The maintaining of PMN homeostasis is an intricate process and the precondition of defense function, which involves activation, adhesion, recruitment, apoptosis and efferocytosis. The regulatory mechanism of PMN homeostasis called neutrophil rheostat, it works through several cytokines and cells. Any factors that break the homeostasis will result in the damage of host immunity,and may relate to the occurrence of periodontitis. Moreover, PMN dysfunction, because of host factors or microorganism factors, is closely related to periodontitis, especially those associated with systemic diseases and gene defect.