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1.
Protein Expr Purif ; 189: 105986, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34600111

RESUMO

To date, there is no functional characterization of EmGGPPS (from Elizabethkingia meningoseptica sp.F2) as enzymes catalyzing GGPP. In this research, maltose-binding protein (MBP), disulfide bond A (DbsA), disulfide bond C (DbsC), and two other small protein tags, GB1 (Protein G B1 domain) and ZZ (Protein A IgG ZZ repeat domain), were used as fusion partners to construct an EmGGPPS fusion expression system. The results indicated that the expression of MBP-EmGGPPS was higher than that of the other four fusion proteins in E. coli BL21 (DE3). Additionally, using EmGGPPS as a catalyst for the production of GGPP was verified using a color complementation assay in Escherichia coli. In parallel with it, the enzyme activity experiment in vitro showed that the EmGGPPS protein could produce GGPP, GPP and FPP. Finally, we successfully demonstrated MK-4 production in engineered E. coli by overexpression of EmGGPPS.

2.
ACS Appl Mater Interfaces ; 13(45): 54439-54446, 2021 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-34738782

RESUMO

Stimuli-responsive liquid crystal elastomers (LCEs), which exhibit sophisticated and versatile shape variations and functions upon stimulations, have constantly interested material science researchers. To date, many challenges still exist in scaling up orientated LCEs with sophisticated physical shapes and multi-functions. Herein, LCEs with various customizable conventional and exotic three-dimensional (3D) shapes and with sizes larger than those previously reported have been prepared by combining magnetic field alignment and soft lithography technology. These LCEs have film, cylinder, ellipsoid, hemispheroid, tube, pyramid, triangle and rectangle frame, grid pattern, cubic frame, and spring shapes. Meanwhile, diversified deformation behaviors such as contraction, expansion, bending, and twisting have been achieved by effectively controlling the alignment directions. Finally, the LCE actuator with hemispheroid shape has been explored for its possible applications in dynamic Braille displays or lenses with adjustable focal length. The simple strategy reported here provides a convenient way to customize multimorphological large-size 3D LCE actuators and their stimuli-responsive deformations. These systems will considerably enlarge the potential applications of LCEs and benefit the development of LCE soft robots and the future special bionic systems.

3.
Bioengineered ; 12(1): 8845-8862, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34606406

RESUMO

Our study aimed to explore potential new diagnostic biomarkers in patients with oral squamous cell carcinoma (OSCC) to find new target molecules involved in the progression of OSCC. Potential novel biomarkers of OSCC were identified using a protein microarray assay. Compared with the healthy control group, there were five proteins (I309, GDF15, AXL, MMP3, and CTACK) in the serum of in situ oral cancer group. However, there were four differentially expressed proteins (MCSF, I309, MMP3, and CTACK) in the serum of the OSCC group. Receiver operating characteristic (ROC) curve analysis results suggested that these six proteins (I309, GDF15, AXL, MMP3, CTACK, and MCSF) had diagnostic value for OSCC. Based on The Cancer Genome Atlas (TCGA) database, we found that only GDF15 expression was associated with the prognosis of OSCC. Subsequently, we verified the expression levels of six proteins in HSC-3 and HaCaT cells, and the results showed that the level of these six proteins was significantly higher in HSC-3 cells than in normal HaCaT cells. Similarly, in the OSCC nude mouse model, the expression levels of these proteins were significantly upregulated in OSCC tumor tissue compared to the normal tissue. GDF15, MMP3, AXL, MCSF, I309, and CTACK may be used as biomarkers for OSCC diagnosis and provide a novel study direction for the treatment of OSCC.

4.
Biotechnol Lett ; 43(12): 2209-2216, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34606014

RESUMO

OBJECTIVES: The production of riboflavin with Bacillus subtilis, is an established process, however it is yet to be fully optimized. The aim of this study was to explore how riboflavin yields can be improved via in vitro and in vivo metabolic engineering modification of the pentose phosphate pathway (PPP). RESULTS: In vitro, glucose was replaced with sodium gluconate to enhance PPP. Flask tests showed that the riboflavin titer increased from 0.64 to 0.87 g/L. The results revealed that the direct use of sodium gluconate could benefit riboflavin production. In vivo, gntP (encoding gluconate permease) was overexpressed to improve sodium gluconate uptake. The riboflavin titer reached 1.00 g/L with the mutant B. subtilis RF01. Ultimately, the fermentation verification of the engineered strain was carried out in a 7-L fermenter, with the increased riboflavin titer validating this approach. CONCLUSIONS: The combination of metabolic engineering modifications in vitro and in vivo was confirmed to promote riboflavin production efficiently by increasing PPP and has great potential for industrial application. This work is aimed to explore how to improve the riboflavin yield by the rational renovation of the pentose phosphate pathway (PPP). In vitro, metabolic engineering mainly uses sodium gluconate as a carbon source instead of glucose, and in vivo, metabolic engineering mainly includes the overexpression of sodium gluconate utility-related genes. The effect of sodium gluconate on cell growth, riboflavin production was investigated in the flasks and fermenter scale.

5.
Sci Rep ; 11(1): 14031, 2021 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-34234236

RESUMO

The screening of non-small cell lung cancer (NSCLC) tumors for anaplastic lymphoma receptor tyrosine kinase (ALK) gene rearrangements is important because of the dramatically favorable therapy response to ALK inhibitor. However, the exact mechanism of poor survival in ALK fusion lung cancer patients without receiving targeted therapy is unclear. In this study, total of 521 tumor specimens from Chinese patients with lung cancer were screened for ALK fusion by immunohistochemistry (IHC) and confirmed by fluorescence in situ hybridization (FISH). As results, there were no cases of coexisting EGFR and ALK mutations identified. Fourteen cases (2.7%) harbored ALK fusion, including eight solid adenocarcinomas with signet ring cell features, four acinar adenocarcinomas with cribriform pattern containing mucin, one adenosquamous carcinoma and one micropapillary adenocarcinoma with mucin. Six (42.9%) of fourteen patients with ALK-positive lung cancer had stage IV disease, and five ALK-positive patients treated with platinum-based chemotherapy had poor outcome (all patients were dead and the mean survival time was 12 months), compared to 72 months for patients with ALK inhibitor therapy. Furthermore, Five ALK-positive cases were analyzed by whole exome sequencing (WES) and via direct transcript counting using a digital probe-base (NanoString) to explore the driver genes. Deregulation of PI3K/AKT signaling pathway in ALK-positive lung cancer was demonstrated by WES analysis, and significantly increased mRNA of ALK, ROS1, MET, SPP1 and PI3K signaling pathway was identified by NanoString assay. The concordance between NanoString, IHC and FISH methodologies for detecting ALK fusion was 100%. Significant overexpression of SPP1 protein in ALK-positive lung cancer was confirmed by IHC compared to paired adjacent normal tissues and ALK-negative cancers. Thus we concluded that SPP1 overexpression is associated with poor outcomes for patients with ALK fusion lung cancer without receiving targeted therapy and PI3K/AKT/SPP1 pathway may become the promising targets in patients with aggressive lung cancer.


Assuntos
Quinase do Linfoma Anaplásico/genética , Expressão Gênica , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/mortalidade , Proteínas de Fusão Oncogênica/genética , Osteopontina/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biomarcadores Tumorais , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Carcinoma Pulmonar de Células não Pequenas/terapia , Gerenciamento Clínico , Receptores ErbB/genética , Feminino , Rearranjo Gênico , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/terapia , Masculino , Pessoa de Meia-Idade , Mutação , Prognóstico , Sequenciamento Completo do Exoma , Adulto Jovem
6.
Int J Gen Med ; 14: 3529-3537, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34290524

RESUMO

Objective: Controversy remains regarding which therapy to initially select for severe aplastic anemia (SAA) patients aged 35-50. This cost-effectiveness analysis aimed to use the Markov model to compare immunosuppressive therapy (IST) with hematopoietic stem-cell transplantation (HSCT) in age-stratified patients with SAA. Methods: A cost-effectiveness analysis using a Markov model compared IST with HSCT in age-stratified patients with SAA. Baseline data were derived from a systematic literature review and collected from Huashan Hospital, Fudan University. The primary outcome was an incremental cost-effectiveness ratio (ICER). Results: The HSCT strategy dominated in patients aged 18-35 even though it was $146,970 more expensive than IST, and the ICER of HSCT to IST was $14,054.19/quality-adjusted life-year (QALY), which was less than the willingness-to-pay value of $25,397.57/QALY. The IST strategy dominated in patients aged 35-50, because it was $72,009 less expensive than HSCT and yielded 3.24 QALYs more than HSCT. The model was vigorous in the sensitivity analyses of the key variables tested through the plausible ranges that were acquired from costing sources and previously published literature. Conclusion: The preferred induction strategy for patients aged 18-35 with SAA appears to be HSCT, and the preferred strategy for patients aged 35-50 is IST, which minimizes costs while maximizing QALYs.

7.
Zhongguo Zhen Jiu ; 41(6): 603-7, 2021 Jun 12.
Artigo em Chinês | MEDLINE | ID: mdl-34085475

RESUMO

OBJECTIVE: To observe the analgesic effect of auricular point sticking therapy during the perioperative stage in the patients with partial lung resection. METHODS: A total of 92 patients with partial lung resection were randomized into an auricular point group (31 cases, 1 case dropped off), the sham-auricular point group (30 cases) and a medication group (31 cases, 1 case dropped off). The routine medication for analgesia was provided in all of the three groups. In the auricular point group, 1 day before operation, the auricular point sticking therapy was applied at shenmen (TF4), sympathetic (AH6a), subcortex (AT4), brain stem (AT3,4i), anterior ear lobe (LO4), chest (AH10) and lung (CO14), retained till the third day after operation. In the sham-auricular point group, the auricular adhesive tape without semen vaccariae was used at the same points as the auricular point group. Separately, the scores of visual analogue scale (VAS) in 8, 16, 24, 48 h and 72 h after operation, the time for obtaining the mean of the postoperative VAS score<3 points, the scores of hospital anxiety and depression scale (HAD) in 8, 72 h after operation and the plasma concentration of ß-endorphin in 24, 48 h after operation, as well as the adverse reactions after operation were recorded in the patients of each group. RESULTS: In 8, 16, 24, 48 h and 72 h after operation, VAS scores in the auricular point group were lower than those in the sham-auricular point group and the medication group separately (P<0.05). In the auricular point group, 16 h after operation the mean of VAS score was less than 3 points, but in the sham-auricular point group and the medication group, 48 h after operation, such a mean score of VAS was obtained. In 8, 72 h after operation, HAD score in the auricular point group was lower than that in the sham-auricular point group and the medication group respectively. In 24, 48 h after operation, the concentration of plasmaß-endorphin was all higher than that in the sham-auricular point group and the medication group respectively (P<0.05). In 24 h after operation, the plasma concentration ofß-endorphin in the sham-auricular point group was higher than the medication group (P<0.05). The incidence of adverse reaction in the auricular point group was lower than that in the sham-auricular point group and the medication group respectively (P<0.05). CONCLUSION: Auricular point sticking therapy relieves perioperative pain, shortens analgesic time, releases anxious and depressive emotions and reduces postoperative adverse reaction in the patients with partial lung resection. The analgesic mechanism is probably related to the increase of plasma concentration of ß-endorphin.


Assuntos
Acupuntura Auricular , Pontos de Acupuntura , Humanos , Pulmão , Dor , Manejo da Dor
8.
Sensors (Basel) ; 21(10)2021 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-34064653

RESUMO

The pattern of bounding box representation and regression has long been dominant in CNN-based pedestrian detectors. Despite the method's success, it cannot accurately represent location, and introduces unnecessary background information, while pedestrian features are mainly located in axis-line areas. Other object representations, such as corner-pairs, are not easy to obtain by regression because the corners are far from the axis-line and are greatly affected by background features. In this paper, we propose a novel detection pattern, named Axis-line Representation and Regression (ALR), for pedestrian detection in road scenes. Specifically, we design a 3-d axis-line representation for pedestrians and use it as the regression target during network training. A line-box transformation method is also proposed to fit the widely used box-annotations. Meanwhile, we explore the influence of deformable convolution base-offset on detection performance and propose a base-offset initialization strategy to further promote the gain brought by ALR. Notably, the proposed ALR pattern can be introduced into both anchor-based and anchor-free frameworks. We validate the effectiveness of ALR on the Caltech-USA and CityPersons datasets. Experimental results show that our approach outperforms the baseline significantly through simple modifications and achieves competitive accuracy with other methods without bells and whistles.


Assuntos
Pedestres , Humanos
9.
Sci Total Environ ; 787: 147644, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34004536

RESUMO

Wetlands provide a broad range of ecosystem services, such as flood control, groundwater replenishment, and water purification. These services are particularly important in the Yellow River Delta, one of four estuarine deltas in China. The aim of this study was to examine the patterns and drivers forces of wetland landscape in the Yellow River Delta. We analyzed the spatio-temporal characteristics of land use change and dynamic landscape patterns changes between 1980 and 2018, divided into eight periods, from land use and remote sensing data. We also analyzed data for annual precipitation, annual temperature, annual evapotranspiration, digital elevation, slope, distance from the main river, built-up land, GDP, and population with principal component analysis, to identify the main drivers behind the changes in the wetland landscape. The results showed that, from 1980 to 2018, the total area of wetland decreased first and then increased the total area of wetland increased and was 1172.73 km2 greater in 2018 than it was in 1980, and the types of wetland in the Yellow River Delta changed frequently. From 1980 to 2005, the area of wetland decreased and the landscape dominance and degree of fragmentation were relatively high. From 2006 to 2018, as environmental policies were implemented and wetland protection became more important, the rate of development of wetland areas increased in a north-south direction, the proportion of landscape types became more balanced, and the spatial distribution homogenized. The main drivers of change in the wetland landscapes were GDP, population, precipitation, and temperature which were included 81.852% of the original information. The findings from this study provide us with an improved understanding of how land use and wetland landscapes changed from 1980 and 2018 and may have implications for the protection of wetland ecosystems, species diversity, and sustainable development.

10.
Front Plant Sci ; 12: 652959, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33897744

RESUMO

Loquat fruit accumulates lignin in its flesh when undergoing chilling injury during postharvest storage, making it a suitable model for the study of flesh lignification. Transcriptional regulation of lignin biosynthesis is principally controlled by the NAC-MYB transcriptional cascade in model plants. Previous research has demonstrated that EjMYB8 activates lignin biosynthesis through direct interaction with the promoter of Ej4CL1. However, the classic NAC-MYB gene regulation network has not been established. Here, the MADS-box gene EjAGL65 was discovered by screening a cDNA library using the EjMYB8 promoter as bait in yeast. A phylogenetic analysis and structural comparisons revealed that EjAGL65 belongs to the Mδ subgroup of the MADS-box family, whose members have not been reported as being involved in the regulation of lignin deposition. EjAGL65 transcription was downregulated at 0°C compared to 5°C, indicating a negative correlation with the change of lignin content. A dual-luciferase assay indicated that EjAGL65 is capable of inhibiting the promoter activity of EjMYB8 in vivo. These results showed that the Mδ MADS-box gene EjAGL65 transcriptionally regulates EjMYB8 during postharvest chilling induced flesh lignification, which differs from the classical regulation model of lignin biosynthesis that has been illustrated for developmental lignin accumulation.

11.
Mol Ther Methods Clin Dev ; 21: 54-66, 2021 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-33768129

RESUMO

Bromodomain protein BRD4 reads histone acetylation (H3K27ac), an epigenomic mark of transcription enhancers. CCAAT enhancer binding protein delta (CEBPD) is a transcription factor typically studied in metabolism. While both are potent effectors and potential therapeutic targets, their relationship was previously unknown. Here we investigated their interplay in vascular smooth muscle cell (SMC) inflammation. Chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) revealed H3K27ac/BRD4 enrichment at Cebpd in injured rat carotid arteries. While genomic deletion of BRD4-associated enhancer in SMCs in vitro decreased Cebpd transcripts, BRD4 gene silencing also diminished Cebpd mRNA and protein, indicative of a BRD4 control over CEBPD expression. Bromodomain-1, but not bromodomain-2, accounted for this BRD4 function. Moreover, endogenous BRD4 protein co-immunoprecipitated with CEBPD, and both proteins co-immunoprecipitated the Cebpd promoter and enhancer DNA fragments. These co-immunoprecipitations (coIPs) were all abolished by the BRD4-bromodomain blocker JQ1, suggesting a BRD4/CEBPD /promoter/enhancer complex. While BRD4 and CEBPD were both upregulated upon tumor necrosis factor alpha (TNF-α) stimulation of SMC inflammation (increased interleukin [IL]-1b, IL-6, and MCP-1), they mediated this stimulation via preferentially elevated expression of platelet-derived growth factor receptor alpha (PDGFRα, versus PDGFRß), as indicated by loss- and gain-of-function experiments. Taken together, our study unravels a hierarchical yet collaborative BRD4/CEBPD relationship, a previously unrecognized mechanism that prompts SMC inflammation and may underlie other pathophysiological processes as well.

12.
JACC Basic Transl Sci ; 6(3): 257-283, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33778212

RESUMO

Polo-like kinase 4 (PLK4) is canonically known for its cytoplasmic function in centriole duplication. Here we show a noncanonical PLK4 function of regulating the transcription factor SRF's nuclear activity and associated myofibroblast-like cell-type transition. In this context, we have further found that PLK4's phosphorylation and transcription are respectively regulated by PDGF receptor and epigenetic factor BRD4. Furthermore, in vivo experiments suggest PLK4 inhibition as a potential approach to mitigating vascular fibrosis.

13.
Environ Pollut ; 274: 116591, 2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33545524

RESUMO

Pharmaceutical antibiotics have recently become emerging environmental contaminants. To enhance the removal efficiency of antibiotics in water, hierarchically porous carbons (HPCs) with designed porous patterns are used in both batch and column mode adsorption processes in this study, and the role of their nanoporous geometry in the adsorption dynamics are explored. THPC (HPC with trimodal pores) and DHPC (HPC with bimodal pores) exhibit remarkably superior adsorption performances to the selected antibiotics than those of commercial activated carbon (AC) with similar surface area, especially in column mode adsorption. The effective treatment volumes of the HPC-columns remain up to 8-10 times those of the AC-columns for the removal of tetracycline and 4-6 times for the removal of tylosin. The mass transfer rates of the carbon-based columns present the order of THPC > DHPC > AC. As comparison, the columns based on monomodal mesoporous carbon (MEC) and microporous carbon (MAC) exhibit low effective treatment volumes although their high mass transfer speed. The interconnected meso/macropores in HPCs benefit the intraparticle mass transfer of guest molecules and the accessibility of adsorption sites. The micropores linking to the meso/macropores not only provide adsorption sites but also facilitate adsorption affinity.


Assuntos
Antibacterianos , Nanoporos , Adsorção , Porosidade , Água
14.
Carbohydr Polym ; 256: 117603, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-33483083

RESUMO

Organic matter pollution and heavy metal pollution have become one of the main problems in water recycling, and the strategy to simultaneously remove soluble organic matter and metal ions is crucial for sewage treatment. In this study, multilayer calcium Alginate beads (n-Alg-DBs-Bas) containing Diatom Biosilica (DBs) and Bacillus subtilis (Bas) were designed as microecologics for sewage treatment. The introduction of DBs in beads and the multilayer structure could promote Bas growth, prolong the stability of the beads, and enhance the adsorption of beads, further improve the sewage treatment efficiency. The organic matter degradation of 3 layered Alg-DBs-Bas reached to 68.23 ± 0.95 % of COD and 58.88 ± 0.84 % of NH4+-N, and the metal ion adsorption was up to 119.31 mg/g for Fe3+, 110.81 mg/g for Zn2+ and 141.34 mg/g for Cu2+. The prepared multilayer calcium alginate beads combined organic matter degradation and metal ions absorption, which is significant for environmental applications.


Assuntos
Alginatos/química , Bacillus subtilis/metabolismo , Diatomáceas/metabolismo , Esgotos , Eliminação de Resíduos Líquidos , Adsorção , Análise da Demanda Biológica de Oxigênio , Cobre/química , Ecologia , Íons , Ferro/química , Cinética , Microscopia Eletrônica de Varredura , Poluentes Químicos da Água/química , Purificação da Água/métodos , Zinco/química
15.
J Nat Prod ; 84(1): 110-119, 2021 01 22.
Artigo em Inglês | MEDLINE | ID: mdl-33356258

RESUMO

Chemical screening of Streptomyces sp. NRRL S-4 with liquid chromatography-mass spectrometry (LC-MS) and the following chromatographic isolation led to the discovery of four 20-membered macrolides, venturicidin A (4) and three new congeners venturicidins D-F (1-3). Genome sequencing of strain S-4 revealed the presence of a biosynthetic gene cluster (BGC) encoding glycosylated type I polyketides (PKS). The BGC designated to venturicidin biosynthesis (ven) was supported by the proposed biosynthetic pathway and confirmed by inactivation of the core PKS gene of venK. Bioinformatic analyses on the conserved motifs and known stereospecificities in PKS modules are consistent with the structure and absolute configuration. This is the first report of venturicidin BGC since the discovery of the macrolide in 1961. In the biological assays, venturicidin A (4) and E (2) displayed a high selective cytotoxicity against acute monocytic leukemia MV-4-11 cells with IC50 values of 0.09 and 0.94 µM, respectively. Venturicidin A (4) also showed a weak inhibitory activity on FMS-like-tyrosine kinase.


Assuntos
Leucemia Monocítica Aguda/genética , Macrolídeos/farmacologia , Streptomyces/química , Venturicidinas/farmacologia , Vias Biossintéticas/genética , Biologia Computacional , Leucemia Monocítica Aguda/metabolismo , Macrolídeos/química , Macrolídeos/isolamento & purificação , Macrolídeos/metabolismo , Família Multigênica , Streptomyces/genética , Venturicidinas/química , Venturicidinas/isolamento & purificação , Venturicidinas/metabolismo
16.
Life Sci Alliance ; 4(1)2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33234676

RESUMO

The sigma-2 receptor (S2R) has long been pharmacologically targeted for antipsychotic treatment and tumor imaging. Only recently was it known for its coding gene and for its role implicated in cholesterol homeostasis. Here, we have investigated the transcriptional control of S2R by the Bromo/ExtraTerminal epigenetic reader family (BETs, including BRD2, 3, and 4) upon cholesterol perturbation. Cholesterol deprivation was induced in ARPE19 cells using a blocker of lysosomal cholesterol export. This condition up-regulated S2R mRNA and protein, and also SREBP2 but not SREBP1, both transcription factors key to cholesterol/fatty acid metabolism. Silencing BRD2 but not BRD3 or BRD4 (though widely deemed a master regulator) averted S2R up-regulation that was induced by cholesterol deprivation. Silencing SREBP2 but not SREBP1 diminished S2R expression. Furthermore, endogenous BRD2 co-immunoprecipitated with the transcription-active N-terminal half of SREBP2, and chromatin immunoprecipitation-qPCR signified co-occupancy of BRD2, H3K27ac (histone acetylation), and SREBP2Nterm at the S2R gene promoter. In summary, this study reveals a previously unrecognized BRD2/SREBP2 cooperative regulation of S2R transcription, thus shedding new light on signaling in response to cholesterol deprivation.


Assuntos
Colesterol/metabolismo , Células Epiteliais/metabolismo , Proteínas Nucleares/metabolismo , Receptores sigma/metabolismo , Transdução de Sinais/genética , Fatores de Transcrição/metabolismo , Androstenos/farmacologia , Anticolesterolemiantes/farmacologia , Azepinas/farmacologia , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Células Epiteliais/efeitos dos fármacos , Regulação da Expressão Gênica , Inativação Gênica , Células HEK293 , Humanos , Proteínas Nucleares/genética , Epitélio Pigmentado da Retina/citologia , Transdução de Sinais/efeitos dos fármacos , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 2/genética , Proteína de Ligação a Elemento Regulador de Esterol 2/metabolismo , Fatores de Transcrição/genética , Transcrição Genética/genética , Transfecção , Triazóis/farmacologia , Regulação para Cima/genética
17.
J Surg Res ; 257: 294-305, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32871430

RESUMO

BACKGROUND: Drug-eluting stents impair post-angioplasty re-endothelialization thus compromising restenosis prevention while heightening thrombotic risks. We recently found that inhibition of protein kinase RNA-like endoplasmic reticulum kinase (PERK) effectively mitigated both restenosis and thrombosis in rodent models. This motivated us to determine how PERK inhibition impacts re-endothelialization. METHODS: Re-endothelialization was evaluated in endothelial-denuded rat carotid arteries after balloon angioplasty and periadventitial administration of PERK inhibitor in a hydrogel. To study whether PERK in smooth muscle cells (SMCs) regulates re-endothelialization by paracrinally influencing endothelial cells (ECs), denuded arteries exposing SMCs were lentiviral-infected to silence PERK; in vitro, the extracellular vesicles isolated from the medium of PDGF-activated, PERK-upregulating human primary SMCs were transferred to human primary ECs. RESULTS: Treatment with PERK inhibitor versus vehicle control accelerated re-endothelialization in denuded arteries. PERK-specific silencing in the denuded arterial wall (mainly SMCs) also enhanced re-endothelialization compared to scrambled shRNA control. In vitro, while medium transfer from PDGF-activated SMCs impaired EC viability and increased the mRNA levels of dysfunctional EC markers, either PERK inhibition or silencing in donor SMCs mitigated these EC changes. Furthermore, CXCL10, a paracrine cytokine detrimental to ECs, was increased by PDGF activation and decreased after PERK inhibition or silencing in SMCs. CONCLUSIONS: Attenuating PERK activity pharmacologically or genetically provides an approach to accelerating post-angioplasty re-endothelialization in rats. The mechanism may involve paracrine factors regulated by PERK in SMCs that impact neighboring ECs. This study rationalizes future development of PERK-targeted endothelium-friendly vascular interventions.


Assuntos
Angioplastia com Balão/efeitos adversos , Reestenose Coronária/prevenção & controle , Miócitos de Músculo Liso/efeitos dos fármacos , Inibidores de Proteínas Quinases/administração & dosagem , Reepitelização/efeitos dos fármacos , eIF-2 Quinase/antagonistas & inibidores , Angioplastia com Balão/instrumentação , Animais , Artérias Carótidas/efeitos dos fármacos , Artérias Carótidas/patologia , Artérias Carótidas/cirurgia , Reestenose Coronária/etiologia , Modelos Animais de Doenças , Stents Farmacológicos/efeitos adversos , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/patologia , Endotélio Vascular/citologia , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/patologia , Humanos , Masculino , Músculo Liso Vascular/citologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Comunicação Parácrina/efeitos dos fármacos , Comunicação Parácrina/genética , RNA Interferente Pequeno/metabolismo , Ratos , Reepitelização/genética , eIF-2 Quinase/genética
18.
Chem Biodivers ; 17(12): e2000769, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33140544

RESUMO

Caryolanes, known as ß-caryophyllene alcohols, are widely occurring sesquiterpenes in plants. From the cultures of marine Streptomyces sp. AH25, two new caryolane sesquiterpenes, micaryolanes A and B (1 and 2), together with caryolan-1,9ß-diol (3) were isolated. Their structures were elucidated by extensive analyses of HR-MS and NMR spectroscopic data. The absolute configurations were assigned via the CD data of the in situ formed [Rh2 (OCOCF3 )4 ] complex and supported by comparison of experimental and calculated specific rotation values. Compounds 1-3 exhibited no activities against Hep3B or MG-63 cell lines or against Gram-positive and Gram-negative bacteria. The results not only enriched the caryolane family, but also proved bacteria as a productive source of terpene metabolites.


Assuntos
Antibacterianos/farmacologia , Biologia Marinha , Sesquiterpenos/isolamento & purificação , Sesquiterpenos/farmacologia , Streptomyces/química , Antibacterianos/isolamento & purificação , Linhagem Celular Tumoral , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana
19.
JAMA Netw Open ; 3(11): e2023644, 2020 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-33136134

RESUMO

Importance: It remains uncertain whether vitamin C routinely used with oral iron supplements is essential for patients with iron deficiency anemia (IDA). Objective: To compare the equivalence and assess the safety of oral iron supplements plus vitamin C or oral iron supplements alone in patients with IDA. Design, Setting, and Participants: This single-center, open-label, equivalence randomized clinical trial was conducted from January 1, 2016, to December 30, 2017, in Huashan Hospital, Fudan University. Adult patients with newly diagnosed IDA were enrolled. Participants were randomly assigned (1:1) to the oral iron supplements plus vitamin C group or the oral iron supplements-only group. Data analysis was performed from March to December 2018. Interventions: Patients were randomized to receive a 100-mg oral iron tablet plus 200 mg of vitamin C or a 100-mg iron tablet alone every 8 hours daily for 3 months. Main Outcomes and Measures: The primary outcome was the change in hemoglobin level from baseline to 2 weeks of treatment, and an equivalence margin of 1 g/dL in hemoglobin was chosen for the demonstration of comparable efficacy. Secondary outcomes included the change in the reticulocyte percentage after 2 weeks of treatment, the increase in hemoglobin level after 4 weeks of treatment, the increase in serum ferritin level after 8 weeks of treatment, and adverse events. Results: Among the 440 randomized patients (220 each in the oral iron supplements plus vitamin C group and iron-only group; 426 women [96.8%]; mean [SD] age, 38.3 [11.7] years), all were assessed for the primary outcome, and 432 (98.2%) completed the trial. From baseline to the 2-week follow-up, the mean (SD) change in hemoglobin level was 2.00 (1.08) g/dL in the oral iron supplements plus vitamin C group and 1.84 (0.97) g/dL in the oral iron supplements-only group (between-group difference, 0.16 g/dL; 95% CI, -0.03 to 0.35 g/dL), thus meeting the criteria for equivalence. The mean (SD) change in serum ferritin level from baseline to 8-week follow-up was 35.75 (11.52) ng/mL in the vitamin C plus iron group and 34.48 (9.50) ng/mL in the iron-only group (between-group difference, 1.27 ng/mL; 95% CI, -0.70 to 3.24 ng/mL; P = .21). There were no significant differences between the 2 groups regarding the rates of adverse events (46 [20.9%] vs 45 [20.5%]; difference, 0.4%; 95% CI, -6.7% to 8.5%; P = .82). No patient withdrew because of adverse events. Conclusions and Relevance: Among patients with IDA, oral iron supplements alone were equivalent to oral iron supplements plus vitamin C in improving hemoglobin recovery and iron absorption. These findings suggest that on-demand vitamin C supplements are not essential to take along with oral iron supplements for patients with IDA. Trial Registration: ClinicalTrials.gov Identifier: NCT02631668.


Assuntos
Anemia Ferropriva/tratamento farmacológico , Ácido Ascórbico/administração & dosagem , Suplementos Nutricionais , Vitaminas/administração & dosagem , Adulto , Quimioterapia Combinada , Feminino , Compostos Ferrosos/administração & dosagem , Compostos Ferrosos/farmacocinética , Hemoglobinas/análise , Humanos , Masculino , Pessoa de Meia-Idade
20.
Atherosclerosis ; 308: 22-31, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32799103

RESUMO

BACKGROUND AND AIMS: Histone methyltransferases are emerging targets for epigenetic therapy. DOT1L (disruptor of telomeric silencing 1-like) is the only known methylation writer at histone 3 lysine 79 (H3K79). It is little explored for intervention of cardiovascular disease. We investigated the role of DOT1L in neointimal hyperplasia (IH), a basic etiology of occlusive vascular diseases. METHODS AND RESULTS: IH was induced via balloon angioplasty in rat carotid arteries. DOT1L and its catalytic products H3K79me2 and H3K79me3 (immunostaining) increased by 4.69 ± 0.34, 2.38 ± 0.052, and 3.07 ± 0.27 fold, respectively, in injured (versus uninjured) carotid arteries at post-injury day 7. Dot1l silencing via shRNA-lentivirus infusion in injured arteries reduced DOT1L, H3K79me2, and IH at day 14 by 54.5%, 37.1%, and 76.5%, respectively. Moreover, perivascular administration of a DOT1L-selective inhibitor (EPZ5676) reduced H3K79me2, H3K79me3, and IH by 56.1%, 58.6%, and 39.9%, respectively. In addition, Dot1l silencing and its inhibition (with EPZ5676) in vivo in injured arteries boosted smooth muscle α-actin immunostaining; pretreatment of smooth muscle cells with EPZ5676 in vitro reduced pro-proliferative marker proteins, including proliferating cell nuclear antigen (PCNA) and cyclin-D1. CONCLUSIONS: While DOT1L is upregulated in angioplasty-injured rat carotid arteries, either its genetic silencing or pharmacological inhibition diminishes injury-induced IH. As such, this study presents a strong rationale for continued mechanistic and translational investigation into DOT1L targeting for treatment of (re)stenotic vascular conditions.


Assuntos
Histona-Lisina N-Metiltransferase , Neointima , Animais , Inativação Gênica , Histona-Lisina N-Metiltransferase/genética , Histona-Lisina N-Metiltransferase/metabolismo , Histonas/metabolismo , Hiperplasia , Ratos
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