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1.
Stem Cells ; 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31574189

RESUMO

Mitochondrial phosphoenolpyruvate carboxykinase (PCK2) is a rate-limiting enzyme that plays critical roles in multiple physiological processes. The decompensation of PCK2 leads to various energy metabolic disorders. However, little is known regarding the effects of PCK2 on osteogenesis by human mesenchymal stem cells (hMSCs). Here, we report a novel function of PCK2 as a positive regulator of MSCs osteogenic differentiation. In addition to its well-known role in anabolism, we demonstrate that PCK2 regulates autophagy. PCK2 deficiency significantly suppressed autophagy, leading to the impairment of osteogenic capacity of MSCs. On the other hand, autophagy was promoted by PCK2 overexpression; this was accompanied by increased osteogenic differentiation of MSCs. Moreover, PCK2 regulated osteogenic differentiation of MSCs via AMP-activated protein kinase (AMPK)/unc-51 like autophagy activating kinase 1(ULK1)-dependent autophagy. Collectively, our present study unveiled a novel role for PCK2 in integrating autophagy and bone formation, providing a potential target for stem cell-based bone tissue engineering that may lead to improved therapies for metabolic bone diseases. Stem Cells 2019.

2.
J Orthop Sci ; 2019 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-31607516

RESUMO

BACKGROUND: The use of 3D-printed scaffolds in repairing bone defects remains unexplored. We aimed to determine whether the duration of electrochemical deposition (ECD) affects the properties of hydroxyapatite (HA) coatings on 3D-printed titanium (TI) scaffolds as well as the corresponding phenotype of MC3T3-E1 cells seeded on these surfaces. METHODS: Five groups of HA-coated TI scaffolds were produced using different durations of ECD (0, 5, 10, 20, and 30 min) and examined under scanning electron microscopy (SEM). MC3T3-E1 cell adhesion to the HA-coated scaffolds and subsequent proliferation and viability were assessed using SEM, DAPI staining, EdU staining, and Alamar Blue assay, respectively. MC3T3-E1 cell expression of osteogenic genes was analyzed by fluorescence RT-PCR. RESULTS: On SEM, longer ECD durations resulted in more compact HA crystals of differing morphology coated onto the TI scaffolds. MC3T3-E1 cell adhesion differed among the five groups (p < 0.05), with the largest number of cells adhered to the scaffolds prepared with 30 min of ECD, followed by the group prepared with 20 min of ECD. However, the ECD duration of 20 min was associated with the highest cell viability and proliferation rate (both p < 0.05) as well as the highest mRNA expression levels of alkaline phosphatase, collagen I, osteocalcin and runt-related transcription factor 2 among the five groups (p < 0.05). CONCLUSIONS: In the fabrication of HA-coated 3D printed TI scaffolds, an ECD duration of 20 min resulted in scaffolds that best promoted MC3T3-E1 cell viability, proliferation and osteogenic gene expression.

3.
Haematologica ; 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31582538

RESUMO

The cellular cytotoxicity of APY0201, a PIKfyve inhibitor, against multiple myeloma was initially identified in an unbiased in vitro chemical library screen. Activity was confirmed in all 25 cell lines tested and 40% of 100 ex vivo patient derived primary samples, and positively correlated with samples harboring trisomies and inversely related to t(11;14). The broad anti- multiple myeloma activity of PIKfyve inhibitors was further demonstrated in confirmatory screens and showed the superior potency of APY0201 when compared to the PIKfyve inhibitors YM201636 and apilimod, with a mid-point EC50 at nanomolar concentrations respectively in 65%, 40%, and 5% of the tested cell lines. An up-regulation of genes in the lysosomal pathway and increased cellular vacuolization was observed in vitro in cell lines following APY0201 treatment however this cellular response did not correlate well with responsiveness. We confirm that PIKfyve inhibition is associated with activation of the transcription factor EB, a master regulator of lysosomal biogenesis and autophagy. Furthermore, we established an assay measuring autophagy as a predictive marker of APY0201 sensitivity. Overall, these findings suggest promising activity of PIKfyve inhibitors secondary to disruption of autophagy in multiple myeloma and a strategy to enrich for likely responders.

4.
Artigo em Inglês | MEDLINE | ID: mdl-31655880

RESUMO

Atherosclerosis is a major cause of mortalities and morbidities worldwide. It is associated with hyperlipidemia and inflammation, and become chronic by triggering metabolites in different metabolic pathways. Disturbance in the human gut microbiota is now considered a critical factor in the atherosclerosis. Trimethylamine-N-oxide (TMAO) attracts attention and is regarded as a vital contributor in the development of atherosclerosis. TMAO is generated from its dietary precursors choline, carnitine, and phosphatidylcholine by gut microbiota into an intermediate compound known as trimethylamine (TMA), which is then oxidized into TMAO by hepatic flavin monooxygenases. The present review focus on advances in TMAO preventing strategies through probiotics, including, modulation of gut microbiome, metabolomics profile, miRNA, or probiotic antagonistic abilities. Furthermore, possible recommendations based on relevant literature have been presented, which could be applied in probiotics and atherosclerosis-preventing strategies.

5.
Wei Sheng Yan Jiu ; 48(4): 583-593, 2019 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-31601339

RESUMO

OBJECTIVE: To understand the differences in the types and levels of antibiotic resistance genes contamination in the dust of air conditioning filters in hospital inpatient wards. METHODS: Wet cotton swabs were used to collect dust samples from air conditioning filters in 19 wards of 7 departments of a third-grade general hospital in Wuhan. The 24 antibiotic resistance genes related to 6 major antibiotics were qualitatively detected by PCR, and 6 typical resistance genes were detected by real-time PCR. RESULTS: Sulfonamides(sulI, sulII), ß-lactams(mecA, blaOXA-51, blaTEM, blaCTX-M, blaSHV, blaKPC, blaNDM-1, blaIMP, blaVIM), aminoglycosides(aac(6᾿-aph(2᾿, aacC2), macrolides(ermA, ermC, ereA), quinolones(qnrA, qnrB, qnrS), a total of five categories of 19 antibiotic resistance genes were detected in the dust of the filter. These include four carbapenem resistance genes(blaNDM-1, blaIMP, blaVIM, blaKPC). The average of absolute content(copies/g) of the six typical resistance genes from high to low was: sulI(1. 06×10~9)>sulII(1. 78×10~8)>blaNDM-1(3. 97×10~7)>aac(6᾿-aph(2᾿(3. 20×10~7)>blaTEM(1. 03×10~7)>aacC2(1. 13×10~6). Among the seven tested departments, traumatic surgery detected up to 18 resistant gene species and 6 typical genes with the highest absolute content. The absolute content of six typical genes in ICU and surgical wards was higher than medical wards. CONCLUSION: A variety of antibiotic resistance genes are detected in the dust of some hospital ward air conditioning filters, suggesting that there may be current or past pollution of resistant bacteria in the relevant environment.

6.
Se Pu ; 37(9): 963-968, 2019 Sep 08.
Artigo em Chinês | MEDLINE | ID: mdl-31642300

RESUMO

A method for the rapid determination of biotoxin (bongkrekic acid) in the Liushenqu was established using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The samples were extracted with methanol by ultrasonication, then the pH was adjusted to 8 using ammonia. After filtration, the extract was purified using an Oasis Max strong anion exchange resin column. The Waters HSS T3 column (100 mm×2.1 mm, 1.8 µm) was used for the UPLC-MS/MS analysis. The mobile phase was acetonitrile-10 mmol/L ammonium formate solution (containing 0.1% (v/v) formic acid). MS analysis was performed using an electrospray ionization (ESI) source in the negative and multiple reaction monitoring (MRM) modes. Under the optimum conditions, the linear range of bongkrekic acid was 0.5-100 µg/L (correlations coefficient (R2)>0.99). The recoveries of the bongkrekic acid were 80.6%-85.3%. The intra-day and inter-day relative standard deviations (RSDs) were in the range of 4.2%-6.8% and 8.2%-13.2%, respectively. The limit of detection (LOD) and limit of quantification (LOQ) were 0.4 µg/kg and 1.2 µg/kg, respectively. The results showed that bongkrekic acid residues were detected in Liushenqu, thereby confirming the supporting role of our method for the risk monitoring of biotoxins in health foods and Chinese herbal medicines. This method is simple, easy, sensitive, and suitable for the determination of the bongkrekic acid residues in Liushenqu.


Assuntos
Ácido Bongcréquico/análise , Contaminação de Medicamentos , Medicamentos de Ervas Chinesas/análise , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas em Tandem
7.
Eur J Pharmacol ; 864: 172717, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31586637

RESUMO

Homocysteine (Hcy) is an independent risk factor in the development of cardiovascular diseases (CVD). Hyperhomocysteinemia (HHcy), induces the injury of vascular endothelial cells via oxidative stress. Oxymatrine (OMT), one of the main components of Sophora flavescens, has displayed anti-inflammatory, anti-oxidant and anti-apoptotic activity. However, the effect of OMT on the Hcy-induced endothelial injury is not clearly defined yet. The aim of this study was to determine the protective effect of OMT on the Hcy-induced endothelial injury and its mechanisms involved. Human umbilical vein endothelial cells (HUVECs) were cultured in vitro. Methyl thiazolyl tetrazolium assay (MTT), fluorescence staining, flow cytometry and western blotting were used in this study. OMT prevented the Hcy-induced toxicity and apoptosis in HUVECs. Moreover, OMT suppressed Hcy-induced increases in reactive oxygen species, lactate dehydrogenase, malondialdehyde levels and increased superoxide dismutase levels. OMT reversed the Hcy-induced decrease in the protein expression of nuclear factor erythroid-2-related factor 2 (Nrf2). In addition, OMT reversed the Hcy-induced apoptosis related biochemical changes such as decreased mitochondrial membrane potential and Bcl-2/Bax protein ratio, and increased protein expression of caspase-9 and caspase-3. Furthermore, OMT elevated the phosphorylation levels of Akt and eNOS, and the formation of nitric oxide (NO) in injured cells. These results suggest that OMT prevents Hcy-induced endothelial injury by regulating mitochondrial-dependent apoptosis and Akt-eNOS-NO signaling pathways concomitantly with accentuation of Nrf2 expression.

8.
Aging (Albany NY) ; 11(19): 8463-8473, 2019 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-31586991

RESUMO

PURPOSE: The aim of this study was to determine the impact of analyzing age as a continuous variable on survival outcomes and treatment selection for extranodal nasal-type NK/T-cell lymphoma. RESULTS: The risk of mortality increased with increasing age, without an apparent cutoff point. Patients' age, as a continuous variable, was independently associated with overall survival after adjustment for covariates. Older early-stage patients were more likely to receive radiotherapy only whereas young-adult advanced-stage patients tended to receive non-anthracycline-based chemotherapy. A decreased risk of mortality with radiotherapy versus chemotherapy only in early-stage patients (HR, 0.347, P < 0.001) or non-anthracycline-based versus anthracycline-based chemotherapy in early-stage (HR, 0.690, P = 0.001) and advanced-stage patients (HR, 0.678, P = 0.045) was maintained in patients of all ages. CONCLUSIONS: These findings support making treatment decisions based on disease-related risk factors rather than dichotomized chronological age. PATIENTS AND METHODS: Data on 2640 patients with extranodal nasal-type NK/T-cell lymphoma from the China Lymphoma Collaborative Group database were analyzed retrospectively. Age as a continuous variable was entered into the Cox regression model using penalized spline analysis to determine the association of age with overall survival (OS) and treatment benefits.

9.
Eur J Neurosci ; 2019 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-31491043

RESUMO

The role of the fibroblast growth factor (FGF) system in depression has received considerable attention in recent years. To understand the role of this system, it is important to identify the specific members of the FGF family that have been implicated and the various mechanisms that they modulated. Here, we review the role of FGFs in depression and integrate evidence from clinical and basic research. These data suggest that changes in the FGF family are involved in depression and possibly in a wider range of psychiatric disorders. We analyse the abnormalities of FGF family members in depression and their roles in modulating depression-related molecules. The role of the FGF family in depression and related disorders needs to be studied in more detail.

10.
Artigo em Inglês | MEDLINE | ID: mdl-31519780

RESUMO

The bioprocessing industry uses recombinant mammalian cell lines to generate therapeutic biologic drugs. To ensure consistent product quality of the therapeutic proteins it is imperative to have a controlled production process. Regulatory agencies and the biotechnology industry consider cell line ″clonal origin″ an important aspect of maintaining process control. Demonstration of clonal origin of the cell substrate, or production cell line, has received considerable attention in the past few years and the industry has improved methods and devised standards to increase the probability and/or assurance of clonal-derivation 1-4. However, older production cell lines developed before the implementation of these methods, herein referred to as ″legacy cell lines″, may not meet current regulatory expectations for demonstration of clonal-derivation. In this article, the members of the IQ Consortium ″Working Group on Clonality″ present our position that the demonstration of process consistency and product comparability of critical quality attributes (CQAs) throughout the development life cycle should be sufficient to approve a license application without additional genetic analysis to support clonal origin, even for legacy cell lines that may not meet current day clonal-derivation standards. With this commentary we discuss advantages and limitations of genetic testing methods to support clonal-derivation of legacy cell lines and wish to promote a mutual understanding with the regulatory authorities regarding their optional use during early drug development, subsequent to IND application and prior to demonstration of product and process consistency at BLA submission.

11.
AAPS PharmSciTech ; 20(8): 312, 2019 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-31529266

RESUMO

To enhance efficiency, convenience, and safety of Parkinson's disease (PD) treatment for geriatric patients, an advanced suspension of Levodopa/Benserazide hydrochloride (LD/BH) has been prepared by cation-exchange resin and used to synchronize sustained release of LD and BH by optimizing coating parameters and prescription. For the purpose, LD and BH were immobilized on the surface of cation-exchange resin, respectively. Based on HPLC results, the cation-exchange resin showed high loading capacity. The studies on drug loading mechanism indicated that both drugs were immobilized by electrostatic interaction rather than physical adsorption. After PEG modification, pretreated drug-resin complexes were coated by emulsion-solvent evaporation method. In order to control drug release in a sustained manner, coating parameters of drug-resin microcapsules were optimized respectively by single-factor analysis. Further, coating prescription of the microcapsules was optimized to synchronize sustained release of LD and BH in vitro by orthogonal design. Utilizing optimal LD-resin microcapsules and BH-resin microcapsules, LD/BH suspension, containing both of them, was prepared by an optimal formulation and characterized by accelerated test and pharmacokinetic study in vivo. The accelerated test confirmed high stability of LD/BH suspension. According to pharmacokinetic results in vivo, in contrast with LD/BH commercial tablets, LD/BH suspensions did not only synchronize sustained release of both drugs but also show good bioequivalence. As LD/BH sustained release suspension can synchronize sustained release of multiple active ingredients by oral administration, the suspension presents promising oral dosage forms for geriatric patients with PD. An advanced Levodopa/Benserazide hydrochloride (LD/BH) suspension, prepared by cation-exchange resin and optimized microencapsulation, synchronizes sustained releases of LD and BH in vivo to benefit Parkinson's disease treatment for geriatric patients.

12.
J Vet Pharmacol Ther ; 2019 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-31529627

RESUMO

Ceftiofur, a third-generation cephalosporin antibiotic, is being extensively used by pet doctors in China. In the current study, the detection method was developed for ceftiofur and its metabolites, desfuroylceftiofur (DCE) and desfuroylceftiofur conjugates (DCEC), in feline plasma. Then, the pharmacokinetics studies were performed following one single intravenous and subcutaneous injection of ceftiofur sodium in cats both at 5 mg/kg body weight (BW) (calculated as pure ceftiofur). Ceftiofur, DCE, and DCEC were extracted from plasma samples, then derivatized and further quantified by high-performance liquid chromatography. The concentrations versus time data were subjected to noncompartmental analysis to obtain the pharmacokinetics parameters. The terminal half-life (t1/2λz ) was calculated as 11.29 ± 1.09 and 10.69 ± 1.31 hr following intravenous and subcutaneous injections, respectively. After intravenous treatment, the total body clearance (Cl) and volume of distribution at steady-state (VSS ) were determined as 14.14 ± 1.09 ml hr-1  kg-1 and 241.71 ± 22.40 ml/kg, respectively. After subcutaneous injection, the peak concentration (Cmax ; 14.99 ± 2.29 µg/ml) was observed at 4.17 ± 0.41 hr, and the absorption half-life (t1/2ka ) and absolute bioavailability (F) were calculated as 2.83 ± 0.46 hr and 82.95%±9.59%, respectively. The pharmacokinetic profiles of ceftiofur sodium and its related metabolites demonstrated their relatively slow, however, good absorption after subcutaneous administration, poor distribution, and slow elimination in cats. Based on the time of drug concentration above the minimum inhibitory concentration (MIC) (T>MIC) calculated in the current study, an intravenous or subcutaneous dose at 5 mg/kg BW of ceftiofur sodium once daily is predicted to be effective for treating feline bacteria with a MIC value of ≤4.0 µg/ml.

13.
J Sex Med ; 16(11): 1696-1707, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31551192

RESUMO

INTRODUCTION: Recently, circular RNA (circRNA) has been proved to occupy a vital pathological position in many diseases by acting as microRNAs sponges. However, the role of circRNA in female sexual dysfunction (FSD), especially in lubrication disorders (LDs), remains unclear. AIM: The aim of this study was to detect circRNA expression in LDs, analyzed target genes, and pathways regulated by the differently expressed circRNAs. METHODS: In this study, next-generation sequencing was first conducted to produce circRNA expression profiles of FSD groups and normal control groups. Furthermore, differences in expression of 6 randomly selected circRNAs were confirmed through real-time quantitative polymerase chain reaction. Kyoto Encyclopedia of Genes and Genomes biological pathway analysis and Gene Ontology showed that immune processes and infection could be involved in the development of FSDs. MAIN OUTCOME MEASURE: CircRNA expression in vaginal epithelial tissue obtained from women with LDs have been detected. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes biological pathway analysis, and circRNA-microRNA interaction predictions were investigated. RESULTS: Totally, 7,746 circRNAs of vaginal epithelial tissue from women of 2 groups were sequenced. Preliminary judgment revealed that there were 73 circRNAs that have significant differential expression, including 53 downregulated circRNAs and 20 upregulated circRNAs. Research results also displayed that the majority of circRNAs has multiple binding sites of microRNAs, including miR-137, which has been reported to be linked to FSD. CLINICAL IMPLICATIONS: We predicted 10 circRNAs paired with hsa-miR-137-5p, but the mechanism of circRNA involvement in disease development remains to be further explored. STRENGTHS & LIMITATIONS: For the first time, the research disclosed the potential pathogenesis of LDs. However, we only analyzed the expression profile of circRNA in FSD, no specific mechanism was further confirmed or proposed. We still have a preliminary understanding, and more research is needed to explore the target of FSD treatment. CONCLUSION: The results suggest that circRNAs have different expression in the FSD groups and play a vital part in the occurrence and development of FSD. Zhang J, Xia H, Zhang A, et al. Circular RNA Expression Profiles in Vaginal Epithelial Tissue of Women With Lubrication Disorders. J Sex Med 2019;16:1696-1707.

15.
Math Biosci Eng ; 16(5): 4122-4134, 2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31499654

RESUMO

In this study, NaNbO3 with average grain size of ~50 nm and KNbO3 with average grain size of ~300 nm nanocrystals are prepared by the water-based citrate precursor sol-gel process. However, the KNbO3 sample exhibits better photocatalytic performance than that of the NaNbO3 sample by Rh B degradation experiment. By Rietveld refinements and piezoelectric displacement measurements, the KNbO3 with the space group of Bmm2 is ferroelectric while the NaNbO3 with the space group of Pbma is antiferroelectric. The polarization-modulated built-in electric fields in the ferroelectric KNbO3 nanoparticles can efficiently enhance the separation of photo-generated charge carries and thus improve the photocatalytic activity. However, there is no internal electric field in the antiferroelectric grain because of the antiparallel spontaneous polarization in the adjacent unit cell. Therefore, KNbO3 exhibits better oxidizing ability of organic dyes than NaNbO3. The ferroelectric KNbO3 nanoparticles exhibit an optimum photocatalytic performance for a complete degradation of Rh B in 100 min under UV-Vis light irradiation with auxiliary ultrasonic excitation. This study demonstrates that the perovskite-type ferroelectric nanocrystals are potentially to design high-performance catalysts for degradation of contaminant.

16.
Gene ; 716: 144031, 2019 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-31377314

RESUMO

Circular RNAs (circRNAs), a novel class of widespread and diverse endogenous RNAs, have been identified as critical regulators of various cancers, including hepatocellular carcinoma (HCC). However, the specific roles of circRNAs in HCC are largely unknown. In this study, we identified a novel circRNA, circ-IGF1R, in HCC tumour tissues and cell lines. Circ-IGF1R levels were found to be significantly upregulated in HCC tissues compared with levels in paired peritumoural tissues. The high expression levels of circ-IGF1R in HCC were associated with tumour size. Moreover, knocking down circ-IGF1R with siRNA significantly attenuated cell proliferation and induced cell apoptosis and cell cycle arrest in vitro. Further investigation revealed that PI3K/AKT signalling pathway activation was involved in the oncogenic functions of circ-IGF1R in HCC. Our study suggests that circ-IGF1R may be a potential target for the prevention and treatment of HCC.


Assuntos
Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , RNA/metabolismo , Apoptose , Carcinoma Hepatocelular/enzimologia , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Ciclo Celular , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Humanos , Neoplasias Hepáticas/enzimologia , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Fosfatidilinositol 3-Quinase/antagonistas & inibidores , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Regulação para Cima
17.
Cancer Lett ; 464: 37-55, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31465841

RESUMO

Long noncoding RNAs (lncRNAs) are defined as RNA transcripts longer than 200 nucleotides that do not encode proteins. LncRNAs have been documented to exhibit aberrant expression in various types of cancer, including prostate cancer. Currently, screening for prostate cancer results in overdiagnosis. The consequent overtreatment of patients with indolent disease in the clinic is due to the lack of appropriately sensitive and specific biomarkers. Thus, the identification of lncRNAs as novel biomarkers and therapeutic targets for prostate cancer is promising. In the present review, we attempt to summarize the current knowledge of lncRNA expression patterns and mechanisms in prostate cancer. In particular, we focus on lncRNAs regulated by the androgen receptor and the specific molecular mechanism of lncRNAs in prostate cancer to provide a potential clinical therapeutic strategy for prostate cancer.

18.
Appl Environ Microbiol ; 85(21)2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31444197

RESUMO

Lactobacillus plantarum is a potential starter and health-promoting probiotic bacterium. Effective, precise, and diverse genome editing of Lactobacillus plantarum without introducing exogenous genes or plasmids is of great importance. In this study, CRISPR/Cas9-assisted double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) recombineering was established in L. plantarum WCFS1 to seamlessly edit the genome, including gene knockouts, insertions, and point mutations. To optimize our editing method, phosphorothioate modification was used to improve the dsDNA insertion, and adenine-specific methyltransferase was used to improve the ssDNA recombination efficiency. These strategies were applied to engineer L. plantarum WCFS1 toward producing N-acetylglucosamine (GlcNAc). nagB was truncated to eliminate the reverse reaction of fructose-6-phosphate (F6P) to glucosamine 6-phosphate (GlcN-6P). Riboswitch replacement and point mutation in glmS1 were introduced to relieve feedback repression. The resulting strain produced 797.3 mg/liter GlcNAc without introducing exogenous genes or plasmids. This strategy may contribute to the available methods for precise and diverse genetic engineering in lactic acid bacteria and boost strain engineering for more applications.IMPORTANCE CRISPR/Cas9-assisted recombineering is restricted in lactic acid bacteria because of the lack of available antibiotics and vectors. In this study, a seamless genome editing method was carried out in Lactobacillus plantarum using CRISPR/Cas9-assisted double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) recombineering, and recombination efficiency was effectively improved by endogenous adenine-specific methyltransferase overexpression. L. plantarum WCFS1 produced 797.3 mg/liter N-acetylglucosamine (GlcNAc) through reinforcement of the GlcNAc pathway, without introducing exogenous genes or plasmids. This seamless editing strategy, combined with the potential exogenous GlcNAc-producing pathway, makes this strain an attractive candidate for industrial use in the future.

19.
Sex Med ; 2019 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-31303464

RESUMO

INTRODUCTION: Female sexual dysfunction (FSD) is a common endocrine disease that impairs the quality of life for many women. The existing therapy strategies still have many disadvantages. It is necessary to explore new pharmacologic treatments that are effective and safe. AIM: The aim of this study was to explore the effects of soy isoflavone (SI) on FSD in mice and the underlying mechanisms. METHODS AND MAIN OUTCOME MEASURES: Laser Doppler flowmetry was used to determine vaginal blood flow. Serum hormone levels and histologic changes of the vagina were analyzed by enzyme-linked immunosorbent assay (ELISA) and by hematoxylin and eosin (H&E) and Masson's trichome staining. The mRNA and protein expression of endothelial nitric oxide synthase (eNOS) was then evaluated by quantitative real-time polymerase chain reaction and western blot assays. RESULTS: Vaginal blood flow was found to be remarkably lower in adult mice, and SI was shown to increase vaginal blood flow in a dose-dependent manner (P < .05). The results of ELISA and H&E and Masson's trichome staining suggest that SI had a positive effect on FSD, as evidenced by the levels of hormones in serum and histologic changes of the vagina, which changed consistently. In addition, the level of eNOS was positively correlated with the concentration of SI, and eNOS inhibitor was able to reverse the improvement in sexual function induced by SI. CONCLUSION: Our study demonstrated that SI could improve sexual function by upregulating the eNOS pathway. Therefore, SI might serve as a promising candidate for the treatment of sexual dysfunction. Zhang J, Zhu Y, Pan L, et al. Soy Isoflavone Improved Female Sexual Dysfunction of Mice Via Endothelial Nitric Oxide Synthase Pathway. Sex Med 2019; XX:XXX-XXX.

20.
Stem Cell Res Ther ; 10(1): 213, 2019 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-31324207

RESUMO

BACKGROUND: As the representative of fenamic acids, an important group of NSAIDs, flufenamic acid (FFA) has been used for anti-inflammation and analgesia in the clinic. Recently, researches have focused on the role of some members of NSAIDs in promoting osteogenesis. However, little attention has been paid to the subgroup of fenamic acids, and it remains unclear whether FFA and other fenamic acids could regulate mesenchymal stem cells' (MSCs) lineage commitment and bone regeneration. METHODS: Here we treated two kinds of human MSCs with FFA at different concentrations in vitro and examined the effect of FFA on osteogenic differentiation of human MSCs. This was followed by heterotopic bone formation assay in nude mice. In addition, ovariectomized and aged mice were used as osteoporotic models to test the effect of FFA on osteoporosis. Besides, activators and inhibitor of nuclear factor-κB (NF-κB) signaling pathway and western blot were used to clarify the mechanism of the promoting effect of low concentration FFA on osteogenesis. RESULTS: Our results indicated that low concentrations of FFA could significantly enhance osteogenic differentiation of human MSCs in vitro, as well as in vivo. In addition, FFA treatment suppressed bone loss in ovariectomized and aged mice. Mechanistically, FFA at low concentrations promoted osteogenesis differentiation of human MSCs by inhibition of the NF-κB signaling pathway. CONCLUSIONS: Collectively, our study suggested that low concentration FFA could be used in bone tissue engineering or osteoporosis by promoting osteogenic differentiation of human MSCs.

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