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1.
Nat Immunol ; 24(11): 1947-1959, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37845489

RESUMO

Age-associated changes in the T cell compartment are well described. However, limitations of current single-modal or bimodal single-cell assays, including flow cytometry, RNA-seq (RNA sequencing) and CITE-seq (cellular indexing of transcriptomes and epitopes by sequencing), have restricted our ability to deconvolve more complex cellular and molecular changes. Here, we profile >300,000 single T cells from healthy children (aged 11-13 years) and older adults (aged 55-65 years) by using the trimodal assay TEA-seq (single-cell analysis of mRNA transcripts, surface protein epitopes and chromatin accessibility), which revealed that molecular programming of T cell subsets shifts toward a more activated basal state with age. Naive CD4+ T cells, considered relatively resistant to aging, exhibited pronounced transcriptional and epigenetic reprogramming. Moreover, we discovered a novel CD8αα+ T cell subset lost with age that is epigenetically poised for rapid effector responses and has distinct inhibitory, costimulatory and tissue-homing properties. Together, these data reveal new insights into age-associated changes in the T cell compartment that may contribute to differential immune responses.


Assuntos
Subpopulações de Linfócitos T , Transcriptoma , Criança , Humanos , Idoso , Envelhecimento/genética , Epitopos/metabolismo , Análise de Célula Única
3.
EMBO Rep ; 24(9): e56562, 2023 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-37492931

RESUMO

Cochlear hair cell loss is a leading cause of deafness in humans. Neighboring supporting cells have some capacity to regenerate hair cells. However, their regenerative potential sharply declines as supporting cells undergo maturation (postnatal day 5 in mice). We recently reported that reactivation of the RNA-binding protein LIN28B restores the hair cell-regenerative potential of P5 cochlear supporting cells. Here, we identify the LIN28B target Trim71 as a novel and equally potent enhancer of supporting cell plasticity. TRIM71 is a critical regulator of stem cell behavior and cell reprogramming; however, its role in cell regeneration is poorly understood. Employing an organoid-based assay, we show that TRIM71 re-expression increases the mitotic and hair cell-forming potential of P5 cochlear supporting cells by facilitating their de-differentiation into progenitor-like cells. Our mechanistic work indicates that TRIM71's RNA-binding activity is essential for such ability, and our transcriptomic analysis identifies gene modules that are linked to TRIM71 and LIN28B-mediated supporting cell reprogramming. Furthermore, our study uncovers that the TRIM71-LIN28B target Hmga2 is essential for supporting cell self-renewal and hair cell formation.


Assuntos
Cóclea , Células Ciliadas Auditivas , Animais , Humanos , Camundongos , Diferenciação Celular/genética , Cóclea/metabolismo , Perfilação da Expressão Gênica , Células Ciliadas Auditivas/metabolismo , Células-Tronco/metabolismo , Proteínas com Motivo Tripartido/genética , Ubiquitina-Proteína Ligases/metabolismo
4.
Proc Natl Acad Sci U S A ; 118(45)2021 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-34732575

RESUMO

Triplex gene editing relies on binding a stable peptide nucleic acid (PNA) sequence to a chromosomal target, which alters the helical structure of DNA to stimulate site-specific recombination with a single-strand DNA (ssDNA) donor template and elicits gene correction. Here, we assessed whether the codelivery of PNA and donor template encapsulated in Poly Lactic-co-Glycolic Acid (PLGA)-based nanoparticles can correct sickle cell disease and x-linked severe combined immunodeficiency. However, through this process we have identified a false-positive PCR artifact due to the intrinsic capability of PNAs to aggregate with ssDNA donor templates. Here, we show that the combination of PNA and donor templates but not either agent alone results in different degrees of aggregation that result in varying but highly reproducible levels of false-positive signal. We have identified this phenomenon in vitro and confirmed that the PNA sequences producing the highest supposed correction in vitro are not active in vivo in both disease models, which highlights the importance of interrogating and eliminating carryover of ssDNA donor templates in assessing various gene editing technologies such as PNA-mediated gene editing.


Assuntos
Edição de Genes/métodos , Anemia Falciforme/genética , Animais , Reações Falso-Positivas , Subunidade gama Comum de Receptores de Interleucina/genética , Camundongos SCID , Técnicas de Sonda Molecular , Ácidos Nucleicos Peptídicos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico
5.
J Med Virol ; 95(2): e28574, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36772841

RESUMO

Human cytomegalovirus (HCMV) preferentially targets neural progenitor cells (NPCs) in congenitally infected fetal brains, inducing neurodevelopmental disorders. While HCMV expresses several microRNAs (miRNAs) during infection, their roles in NPC infection are unclear. Here, we characterized expression of cellular and viral miRNAs in HCMV-infected NPCs during early infection by microarray and identified seven differentially expressed cellular miRNAs and six significantly upregulated HCMV miRNAs. Deep learning approaches were used to identify potential targets of significantly upregulated HCMV miRNAs against differentially expressed cellular messenger RNA (mRNAs), and the associations with miRNA-mRNA expression changes were observed. Gene ontology enrichment analysis indicated cellular gene targets were significantly enriched in pathways involved in neurodevelopment and cell-cycle processes. Viral modulation of selected miRNAs and cellular gene targets involved in neurodevelopmental processes were further validated by real-time quantitative reverse transcription polymerase chain reaction. Finally, a predicted 3' untranslated region target site of hcmv-miR-US25-1 in Jag1, a factor important for neurogenesis, was confirmed by mutagenesis. Reduction of Jag1 RNA and protein levels in NPCs was observed in response to transient expression of hcmv-miR-US25-1. A hcmv-miR-US25-1 mutant virus (ΔmiR-US25) displayed limited ability to downregulate Jag1 mRNA levels and protein levels during the early infection stage compared with the wild type virus. Our collective experimental and computational investigation of miRNAs and cellular mRNAs expression in HCMV-infected NPCs yields new insights into the roles of viral miRNAs in regulating NPC fate and their contributions to HCMV neuropathogenesis.


Assuntos
Infecções por Citomegalovirus , MicroRNAs , Humanos , MicroRNAs/genética , Citomegalovirus/genética , Células-Tronco/metabolismo
6.
Bioorg Med Chem ; 82: 117234, 2023 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-36906964

RESUMO

Cancer with low survival rates is the second main cause of death among all diseases in the world and consequently, effective antineoplastic agents are urgently needed. Allosecurinine is a plant-derived indolicidine securinega alkaloid shown bioactivity. The object of this study is to investigate synthetic allosecurinine derivatives with considerable anticancer capacity against nine human cancer cell lines as well as mechanism of action. We synthesized twenty-three novel allosecurinine derivatives and evaluated their antitumor activity against nine cancer cell lines for 72 h by MTT and CCK8 assays. FCM was applied to analyze the apoptosis, mitochondrial membrane potential, DNA content, ROS production, CD11b expression. Western blot was selected to analyze the protein expression. Structure-activity relationships were established and potential anticancer lead BA-3 which induced differentiation of leukemia cells towards granulocytosis at low concentration and apoptosis at high concentration was identified. Mechanism studies showed that mitochondrial pathway mediated apoptosis within cancer cells with cell cycle blocking was induced by BA-3. In addition, western blot assays revealed that BA-3 induced expression of the proapoptotic factor Bax, p21 and reduced the levels of antiapoptotic protein such as Bcl-2, XIAP, YAP1, PARP, STAT3, p-STAT3, and c-Myc. Collectively, BA-3 was a lead compound for oncotherapy at least in part, through the STAT3 pathway. These results were an important step in further studies on allosecurinine-based antitumor agent development.


Assuntos
Alcaloides , Antineoplásicos , Compostos Heterocíclicos de Anel em Ponte , Neoplasias , Humanos , Antineoplásicos/farmacologia , Azepinas/farmacologia , Compostos Heterocíclicos de Anel em Ponte/farmacologia , Lactonas/farmacologia , Apoptose , Alcaloides/farmacologia , Proliferação de Células , Ensaios de Seleção de Medicamentos Antitumorais , Linhagem Celular Tumoral
7.
Proc Natl Acad Sci U S A ; 117(36): 22225-22236, 2020 09 08.
Artigo em Inglês | MEDLINE | ID: mdl-32826333

RESUMO

Mechano-sensory hair cells within the inner ear cochlea are essential for the detection of sound. In mammals, cochlear hair cells are only produced during development and their loss, due to disease or trauma, is a leading cause of deafness. In the immature cochlea, prior to the onset of hearing, hair cell loss stimulates neighboring supporting cells to act as hair cell progenitors and produce new hair cells. However, for reasons unknown, such regenerative capacity (plasticity) is lost once supporting cells undergo maturation. Here, we demonstrate that the RNA binding protein LIN28B plays an important role in the production of hair cells by supporting cells and provide evidence that the developmental drop in supporting cell plasticity in the mammalian cochlea is, at least in part, a product of declining LIN28B-mammalian target of rapamycin (mTOR) activity. Employing murine cochlear organoid and explant cultures to model mitotic and nonmitotic mechanisms of hair cell generation, we show that loss of LIN28B function, due to its conditional deletion, or due to overexpression of the antagonistic miRNA let-7g, suppressed Akt-mTOR complex 1 (mTORC1) activity and renders young, immature supporting cells incapable of generating hair cells. Conversely, we found that LIN28B overexpression increased Akt-mTORC1 activity and allowed supporting cells that were undergoing maturation to de-differentiate into progenitor-like cells and to produce hair cells via mitotic and nonmitotic mechanisms. Finally, using the mTORC1 inhibitor rapamycin, we demonstrate that LIN28B promotes supporting cell plasticity in an mTORC1-dependent manner.


Assuntos
Células Ciliadas Auditivas/fisiologia , Células Labirínticas de Suporte/metabolismo , MicroRNAs/metabolismo , Proteínas de Ligação a RNA/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Animais , Regulação da Expressão Gênica no Desenvolvimento , Genótipo , Camundongos , MicroRNAs/genética , Organoides , Proteínas de Ligação a RNA/genética , Serina-Treonina Quinases TOR/genética
8.
Phys Chem Chem Phys ; 25(1): 529-539, 2022 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-36477271

RESUMO

Simulated photoelectron spectroscopy was conducted to investigate the structural evolution and electronic properties of TiASil (A = Sc, Ti; l ≤ 12) clusters and their anions via the Perdew-Burke-Enzerhof scheme and extensive cluster search using the ABCluster software. The results revealed that the ground-state structures of the TiASil (A = Sc, Ti) clusters generally exhibited similar configurations except for the Ti2Si3, ScTiSi3, and TiScSi10 clusters. Furthermore, the TiASil clusters exhibited an adsorptive evolution pattern, and the TiASi4 unit was considered the basic constituent framework of the structure, excluding several distortions and minor changes. With the increase in the cluster size, the lowest-energy structures varied from the exohedral to the cage structures of the single-metal atom at the center. Regarding the second energy difference data, the neutral TiASi4 clusters exhibited better stability among the neutral and anionic TiASil (A = Sc, Ti; l ≤ 12) clusters. Furthermore, the chemical bonding in the TiASi4 clusters was analyzed by molecular orbitals (MOs), highest occupied MO-lowest unoccupied MO gaps, and adaptive natural density partitioning analyses for the best Ti2Si4 cluster especially, and the results were combined with the natural population analysis data. The hybridization of the spd orbital of the metal atoms, eight localized bonds, and four delocalized bonds may primarily account for the relative stabilities of the neutral, square bipyramidal structure of Ti2Si4. Thus, the TiASi4 clusters may be assembled as the basic units of silicon-based semiconductor clusters of large-size neutral and anionic TiASil.

9.
Gastroenterology ; 159(1): 96-104, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32165208

RESUMO

BACKGROUND & AIMS: Biomarkers are needed to identify patients at risk for development of inflammatory bowel diseases. We aimed to identify serum biomarkers of Crohn's disease and ulcerative colitis that can be detected and quantified before diagnosis. METHODS: We obtained serum samples from patients archived before a diagnosis of Crohn's disease (n = 200) or ulcerative colitis (n = 199), as well as from 200 healthy individuals (controls), collected from 1998 through 2013 as part of the US Defense Medical Surveillance System. We measured levels of antibodies against microbes (anti-Saccharomyces cerevisiae IgA or IgG, anti-Escherichiacoli outer membrane porin C, anti-CBir1, anti-flagellin 2, anti-flagellin X, and perinuclear anti-neutrophil cytoplasmic antibodies) and 1129 proteins in each sample. We then used functional principal component analysis to derive the time-varying trajectory for each marker, which then was used in a multivariate model to predict disease status. Predictive performances at different prediagnosis timepoints were evaluated using area under the receiver operating characteristic curves (AUROCs). Biological pathways that were up-regulated in serum from patients with Crohn's disease were identified based on changes in protein abundance at different time periods preceding diagnosis. RESULTS: We identified a panel of 51 protein biomarkers that were predictive of Crohn's disease within 5 years with an AUROC of 0.76 and a diagnosis within 1 year with an AUROC of 0.87. Based on the proteins included in the panel, imminent development of CD was associated with changes in the complement cascade, lysosomes, innate immune response, and glycosaminoglycan metabolism. Serum antibodies and proteins identified patients who received a diagnosis of ulcerative colitis within 5 years with an AUROC of only 0.56 and within 1 year with an AUROC of 0.72. CONCLUSIONS: We identified a panel of serum antibodies and proteins that were predictive of patients who will receive a diagnosis of Crohn's disease within 5 years with high accuracy. By contrast we did not identify biomarkers associated with future diagnosis of ulcerative colitis.


Assuntos
Anticorpos Anticitoplasma de Neutrófilos/sangue , Anticorpos Antibacterianos/sangue , Anticorpos Antifúngicos/sangue , Colite Ulcerativa/diagnóstico , Doença de Crohn/diagnóstico , Adulto , Anticorpos Anticitoplasma de Neutrófilos/imunologia , Anticorpos Antibacterianos/imunologia , Anticorpos Antifúngicos/imunologia , Biomarcadores/sangue , Estudos de Casos e Controles , Colite Ulcerativa/sangue , Colite Ulcerativa/imunologia , Doença de Crohn/sangue , Doença de Crohn/imunologia , Escherichia coli/imunologia , Feminino , Voluntários Saudáveis , Humanos , Imunidade Inata , Masculino , Modelos Estatísticos , Valor Preditivo dos Testes , Prognóstico , Proteômica , Curva ROC , Saccharomyces cerevisiae/imunologia , Fatores de Tempo , Adulto Jovem
10.
Gastroenterology ; 158(3): 515-526.e10, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31711925

RESUMO

BACKGROUND & AIMS: Noninvasive tests to measure endoscopic activity in patients with Crohn's disease (CD) have limitations. We aimed to develop a test to identify patients in remission, based on endoscopic analysis, and monitor CD activity based on serum levels of proteins. METHODS: We developed a test to measure 13 proteins in blood (ANG1, ANG2, CRP, SAA1, IL7, EMMPRIN, MMP1, MMP2, MMP3, MMP9, TGFA, CEACAM1, and VCAM1), called the endoscopic healing index [EHI], using samples from 278 patients with CD from a multinational training cohort. We validated the test using 2 independent cohorts of patients with CD: 116 biologic-naive patients with early-stage CD (validation cohort 1) and 195 biologic-exposed patients with chronic CD (validation cohort 2). The ability of the test to identify patients with active disease vs patients in remission (defined as a simple endoscopic score for CD of ≤2 and ≤1 in each segment, or a total CD endoscopic index of severity score <3) was assessed by using area under receiver operating characteristic curve (AUROC) analysis. The diagnostic accuracy of the test was compared with that of measurement of serum C-reactive protein (CRP) and fecal calprotectin. RESULTS: The EHI scores range from 0 to 100 units; higher scores indicate more severe CD activity, based on endoscopy findings. The EHI identified patients in remission with an AUROC of 0.962 in validation cohort 1 (95% confidence interval, 0.942-0.982) and an AUROC of 0.693 in validation cohort 2 (95% confidence interval, 0.619-0.767), regardless of CD location or phenotype. A cutoff value of 20 points identified patients in remission with the highest level of sensitivity (97.1% in validation cohort 1 and 83.2% in validation cohort 2), with specificity values of 69.0% and 36.6%, respectively. A cutoff value of 50 points identified patients in remission with the highest level of specificity (100% in validation cohort 1 and 87.8% in validation cohort 2), with sensitivity values of 37.3% and 30.0%, respectively. The EHI identified patients in remission with a significantly higher AUROC value than the test for CRP (0.876, P < .001 in validation cohort 1 and 0.624, P = .109 in validation cohort 2). In analysis of patients with available FC measurements, the AUROC value for the EHI did not differ significantly from that of measurement of FC (AUROC, 0.950 for EHI vs AUROC, 0.923 for FC; P = .147 in validation cohort 1 and AUROC, 0.803 for EHI vs AUROC, 0.854 for FC; P = .298 in validation cohort 2). CONCLUSIONS: We developed an index called the EHI to identify patients with CD in endoscopic remission based on blood levels of 13 proteins. The EHI identified patients with resolution of endoscopic disease activity, with good overall accuracy, although with variation between the 2 cohorts assessed. The EHI AUROC values were comparable to measurement of FC and higher than measurement of serum CRP. The test might be used in practice to assess endoscopic activity in patients with CD.


Assuntos
Colonoscopia , Doença de Crohn/diagnóstico , Fármacos Gastrointestinais/administração & dosagem , Infliximab/administração & dosagem , Índice de Gravidade de Doença , Adulto , Biomarcadores/sangue , Proteína C-Reativa/análise , Colo/diagnóstico por imagem , Colo/efeitos dos fármacos , Colo/patologia , Doença de Crohn/sangue , Doença de Crohn/tratamento farmacológico , Fezes/química , Feminino , Humanos , Mucosa Intestinal/diagnóstico por imagem , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/patologia , Complexo Antígeno L1 Leucocitário/análise , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Curva ROC , Recidiva , Indução de Remissão/métodos , Estudos Retrospectivos , Resultado do Tratamento , Adulto Jovem
11.
J Org Chem ; 86(8): 5506-5517, 2021 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-33797258

RESUMO

A practical and efficient protocol toward fully substituted isothiazolones through Selectfluor-mediated intramolecular oxidative annulation of α-carbamoyl ketene dithioacetals has been developed in the presence of H2O and metal-free conditions. Notably, the experimental results reveal that H2O was crucial to the formation of new N-S bonds and the elimination of alkyl group from the sulfur atom. This protocol provides readily prepared substrates and possesses good functional group tolerance, mild reaction conditions, and operational simplicity, which provides potential access to applications in the pharmaceutical chemistry.


Assuntos
Etilenos , Água , Compostos de Diazônio , Cetonas , Estrutura Molecular
12.
Phys Chem Chem Phys ; 23(45): 25679-25688, 2021 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-34755155

RESUMO

The growth patterns, simulated photoelectron spectra, and electronic properties of LaASil (A = Sc, Y, and La; l ≤ 10) compounds and their anions were studied via quantum chemistry calculations using the Perdew-Burke-Ernzerhof (PBE) method and unprejudiced structural searching software ABCluster. The results revealed that the growth patterns of the most stable structures of neutral and anionic LaASil showed an adsorptive mode. The lowest-energy structures (LESs) of the LaASil (l ≤ 7) clusters were similar, except for those of anionic LaYSi4- and LaYSi5- and neutral LaScSi7. Additionally, we investigated and calculated the photoelectron spectra, vertical detachment energies, adiabatic electron affinities, relative stability, charge transfer, magnetic moment, and chemical bond analysis of the LaASil ground-state structures. The La2Sil clusters exhibited higher stability than the LaYSil and LaScSil systems owing to their higher dissociation energies (DEs). The DEs of the LESs in the LaASi3 molecule are higher than those of other clusters. Thus, the LaASi3 cluster shows potential as a building framework for Si-based cluster materials with good stability. The natural population analysis data and chemical bond analysis results showed that the spd hybridization of the orbitals of the metal atoms in the LaASil system occurred. Except for the LaScSi9 and LaScSi10 clusters, the neutral LaASil compounds transform into the corresponding anions when an extra electron is accepted by the Si clusters.

13.
J Clin Periodontol ; 48(10): 1293-1301, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34219259

RESUMO

AIM: To evaluate the outcomes of an apically repositioned flap (ARF) plus xenogeneic collagen matrix (XCM) in augmenting keratinized mucosa width (KMW) around dental implants when compared with ARF plus free gingival grafts (FGG). MATERIALS AND METHODS: Twenty-six participants with at least one site with KMW ≤2 mm were randomized into FGG or XCM group. Clinical examinations were performed at baseline and at 2 and 6 months after surgery, including KMW, keratinized mucosa thickness, gingival index (GI), and probing depth (PD). Post-operative pain and patient satisfaction were also evaluated. RESULTS: At 6 months, FGG attained a greater increase of KMW and thicker mucosa than XCM (4.1 ± 1.6 mm vs. 1.8 ± 1.0 mm, p < .001; 1.7 ± 0.6 mm vs. 1.2 ± 0.3 mm, p < .01). Regarding GI, PD, post-operative pain, aesthetic outcomes, and patient satisfaction, no significant difference could be detected. Moreover, the operation time of XCM group was shorter (60 ± 9 min vs. 39 ± 8 min, p < .001). CONCLUSIONS: FGG could result in greater increase of KMW than XCM, though both could increase KMW, maintain peri-implant health, and attain comparable aesthetic outcomes. The use of XCM was associated with reduced operation time.


Assuntos
Implantes Dentários , Colágeno , Estética Dentária , Gengiva , Gengivoplastia , Humanos , Mucosa
14.
Molecules ; 26(18)2021 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-34576982

RESUMO

Chemical investigation of the Antarctic lichen-derived fungal strain Acremonium sp. SF-7394 yielded a new amphilectane-type diterpene, acrepseudoterin (1), and a new acorane-type sesquiterpene glycoside, isocordycepoloside A (2). In addition, three known fungal metabolites, (-)-ternatin (3), [D-Leu]-ternatin (4), and pseurotin A (5), were isolated from the EtOAc extract of the fungal strain. Their structures were mainly elucidated by analyzing their NMR and MS data. The absolute configuration of 1 was proposed by electronic circular dichroism calculations, and the absolute configuration of the sugar unit in 2 was determined by a chemical method. The inhibitory effects of the isolated compounds on protein tyrosine phosphatase 1B (PTP1B) were evaluated by enzymatic assays; results indicated that acrepseudoterin (1) and [D-Leu]-ternatin (4) dose-dependently inhibited the enzyme activity with IC50 values of 22.8 ± 1.1 µM and 14.8 ± 0.3 µM, respectively. Moreover, compound 1 was identified as a competitive inhibitor of PTP1B.


Assuntos
Acremonium , Proteína Tirosina Fosfatase não Receptora Tipo 1 , Inibidores Enzimáticos
15.
Molecules ; 26(8)2021 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-33921386

RESUMO

Acanthopanax henryi (Oliv.) Harms (Araliaceae), also known as Eleutherococcus henryi and Caoyewujia (Hengliwujia) in Chinese, is a widely used traditional Chinese herb with the effects of expelling wind and removing dampness, relaxing the muscles and stimulating the blood circulation, and regulating the flow of qi to alleviate pain in the theory of Traditional Chinese Medicine. Acanthopanax henryi (AH, thereafter) possesses ginseng-like activities and is known as ginseng-like herb. In the past decade, a great number of phytochemical and pharmacological studies on AH have been carried out. Several kinds of chemical compositions have been reported, including terpenoids (monoterpenoids, diterpenoids, and triterpenoid saponins), phenylpropanoids, caffeoyl quinic acid derivatives, flavonoids, lignans, sterols, fatty acids, etc., among which, triterpenoid saponins were considered to be the most active components. Considerable pharmacological experiments in vitro have demonstrated that AH possessed anti-neuroinflammatory, anti-adipogenic, anti-inflammatory, antibacterial, anti-cancer, anti-oxidation, anti-AChE, anti-BuChE, and antihyaluronidase activities. The present review is an up-to-date and comprehensive analysis of the botany, phytochemistry, and pharmacology of AH.


Assuntos
Eleutherococcus/química , Etnofarmacologia , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/química , Compostos Fitoquímicos/isolamento & purificação , Pesquisa
16.
J Cell Physiol ; 235(10): 7273-7282, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32039486

RESUMO

microRNAs may function as oncogenes or tumor suppressor genes that play crucial roles in human carcinogenesis and cancer development. Growing evidence revealed that the tumor suppressor Id3 is involved in tumor progression, carcinogenesis, and the tumor microenvironment. We identified miR-212-5p as a negative posttranscriptional modulator of Id3. Dual luciferase reporter assay was used to verify that Id3 is a direct target gene of miR-212-5p. Id3 was lowly expressed and miR-212-5p was highly expressed in non-small-cell lung cancer (NSCLC) tissues and cells. In addition, we found that NSCLC patients having a higher level of miR-212-5p expression had a shorter survival time. Besides this, miR-212-5p could directly target Id3 and reduce its expression. miR-212-5p overexpression significantly accelerated cell proliferation, migration, and invasion by reversing the effects of Id3. Id3 overexpression by silencing miR-212-5p expression suppressed phosphatidylinositol 3 kinase (PI3K)/Akt activity and consequently promoted apoptosis and inhibited cell proliferation in lung cancer cells. Consistent with the in vitro results, a xenograft mouse model was used to validate the fact that miR-212-5p could promote tumorigenesis by targeting Id3 and activate the PI3K/Akt pathway in vivo as well. Taken together, the present results indicated that miR-212-5p may be involved in progression of NSCLC through the PI3K/Akt signaling pathway by targeting Id3.


Assuntos
Adenocarcinoma de Pulmão/genética , Adenocarcinoma de Pulmão/metabolismo , Proteínas Inibidoras de Diferenciação/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Proteínas de Neoplasias/metabolismo , Adenocarcinoma de Pulmão/patologia , Animais , Apoptose , Carcinogênese/genética , Carcinogênese/metabolismo , Carcinogênese/patologia , Carcinógenos/metabolismo , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Xenoenxertos , Humanos , Neoplasias Pulmonares/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Invasividade Neoplásica , Fosfatidilinositol 3-Quinases/metabolismo , Prognóstico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais
17.
Cytogenet Genome Res ; 160(5): 238-244, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32659759

RESUMO

X-linked Alport syndrome (XLAS) is a common hereditary nephropathy caused by COL4A5 gene mutations. To date, many splice site mutations have been described but few have been functionally analyzed to verify the exact splicing effects that contribute to disease pathogenesis. Here, we accidentally discovered 2 COL4A5 gene splicing mutations affecting the same residue (c.2917+1G>A and c.2917+1G>C) in 2 unrelated Chinese families. In vitro minigene assays showed that the 2 mutations produced 3 transcripts in H293T cells: one with a 96-bp deletion in exon 33, one with exon 33 skipping, and one with exon 33-34 skipping. However, fragment analysis results showed that the main splicing effects of the 2 mutations were different, the c.2917+1G>A mutation mainly activated a cryptic donor splice site in exon 33 and resulted in the deletion of 96 bp in exon 33, while the c.2917+1G>C mutation mainly caused exon 33 skipping. Our findings indicate that different nucleotide substitutions at the same residue can cause different splicing effects, which may contribute to the variable phenotype of Alport syndrome.


Assuntos
Processamento Alternativo/genética , Povo Asiático/genética , Colágeno Tipo IV/genética , Mutação , Nefrite Hereditária/genética , Sítios de Splice de RNA/genética , Adulto , Linhagem Celular , Criança , Pré-Escolar , Simulação por Computador , Éxons/genética , Feminino , Hematúria/genética , Humanos , Masculino , Linhagem , Proteinúria/genética
18.
Int J Syst Evol Microbiol ; 70(10): 5271-5279, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32833615

RESUMO

A psychrotolerant actinobacterium, designated strain J5903T, was isolated from an alkaline soil sample from the rhizosphere of Suaeda salsa collected in desertification land surrounding Jiuliancheng Nur in Hebei Province, PR China. Cells of the isolate were Gram-stain-positive, aerobic, non-motile and non-spore-forming cocci. Strain J5903T grew optimally at 20‒25 °C, at pH 7.0‒7.5 and with <1 % (w/v) NaCl. The cell-wall peptidoglycan type was B2γ with d-2,4-diaminobutyric acid and l-2,4-diaminobutyric acid as diagnostic amino acids. The muramyl residue was acetyl type. The menaquinones were MK-11, MK-12, MK-10 and MK-13. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and one unidentified glycolipid. The major whole-cell fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. The genomic DNA G+C content was 69.1 mol%. It shared the highest average nucleotide identity and digital DNA-DNA hybridization values with Planctomonas deserti 13S1-3T. Phylogenies based on genome sequence showed that strain J5903T and P. deserti 13S1-3T formed a robust cluster with high bootstrap support. Strain J5903T shared typical chemotaxonomic characteristics with P. deserti 13S1-3T. Combining the polyphasic taxonomic evidence, strain J5903T represents a novel species of the genus Planctomonas, for which the name Planctomonas psychrotolerans sp. nov. is proposed. The type strain is J5903T (=DSM 101894T=CGMCC 1.15523T).


Assuntos
Actinobacteria/classificação , Chenopodiaceae/microbiologia , Filogenia , Rizosfera , Microbiologia do Solo , Actinobacteria/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Glicolipídeos/química , Hibridização de Ácido Nucleico , Peptidoglicano/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/química
19.
Int J Syst Evol Microbiol ; 70(7): 4179-4185, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32558636

RESUMO

A novel moderately halophilic, filamentous actinobacterium, designated as XMNu-373T, was isolated from a saline-alkaline soil sample collected from the Mongolia Plateau, Dongwu County, Inner Mongolia Autonomous Region, PR China. The isolate grew optimally at 28‒37 °C, pH 7.0‒8.0 and with 2-5 % (w/v) NaCl. The substrate mycelia fragmented into rod-like elements, and the white aerial mycelia formed spore chains at maturity. The predominant menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, three unidentified phosphoglycolipids, an unidentified aminophospholipid, two phosphatidylinositol mannosides, four unidentified phospholipids, phosphatidylglycerol and two unidentified lipids. The major cellular fatty acids were iso-C16 : 0, anteiso-C17 : 0 and anteiso-C15 : 0. The genomic DNA G+C content was 66.2 mol%. It shared high 16S rRNA gene sequence similarities to Phytoactinopolyspora halotolerans YIM 96448T (96.1 %) and Phytoactinopolyspora endophytica EGI 60009T (96.0 %). Phylogenetic trees based on 16S rRNA gene sequences revealed that strain XMNu-373T resided in the clade of family Jiangellaceae, and it formed a monophyletic branch distinct from four other recognized type species in the subclade of the genus Phytoactinopolyspora. On the basis of polyphasic taxonomic evidence, strain XMNu-373T represents a novel species of the genus Phytoactinopolyspora, for which the name Phytoactinopolyspora mesophila sp. nov. is proposed. The type strain is XMNu-373T (=JCM 33740T=CGMCC 4.7654T).


Assuntos
Actinobacteria/classificação , Álcalis , Filogenia , Salinidade , Microbiologia do Solo , Actinobacteria/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Solo/química , Vitamina K 2/análogos & derivados , Vitamina K 2/química
20.
J Clin Pediatr Dent ; 44(6): 436-441, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-33378465

RESUMO

BACKGROUND: Plasma cell granuloma (PCG) is a rare benign pseudotumorous proliferation of unclear etiology that is mainly situated in the lungs. Gingival PCG is an even more peculiar lesion that usually occurs in middle-aged or elderly individuals and clinically manifests as a solitary entity. CASE REPORT: A 15-year-old male with no underlying medical conditions presented with multiple gingival masses in the right maxilla, which were initially thought to be epulis. The lesions were resected completely and the excisional biopsies sent for histological examination. Immunohistochemical (IHC) stain revealed dense polyclonal plasma cell infiltration with positive expression of both kappa and lambda light chains, confirming a diagnosis of gingival PCG. Subsequently, the affected gingiva healed uneventfully, with no sign of recurrence over 2 years of follow-up. CONCLUSIONS: The present report depicts an extremely unusual case of gingival PCG occurring in a juvenile with multiple lesions, which is worth attention in clinical pediatric dentistry. Excisional-biopsy and histological investigations are imperative for a confirmative diagnosis and to exclude potential aggressive conditions. Complete resection of lesions seems to be a valid treatment, while long-term clinical follow-up is still needed.


Assuntos
Doenças da Gengiva , Granuloma de Células Plasmáticas , Adolescente , Idoso , Criança , Gengiva , Doenças da Gengiva/diagnóstico , Doenças da Gengiva/cirurgia , Granuloma de Células Plasmáticas/diagnóstico , Granuloma de Células Plasmáticas/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade
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