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1.
Cell Prolif ; : e12694, 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31568642

RESUMO

OBJECTIVES: Mesenchymal stem cell (MSC)-based dental tissue regeneration is a potential treatment method in future, while inflammation and hypoxia niche will affect MSC-mediated tissue regeneration. In this research, we intended to investigate the influence and mechanism of secreted frizzled-related protein 2(SFRP2) on MSC function under inflammation and hypoxia conditions. MATERIAL AND METHODS: Stem cells from apical papilla (SCAPs) were used in this study. The alkaline phosphatase (ALP) activity, Alizarin Red S staining, scratch-simulated wound migration and transwell chemotaxis assay were used to evaluate the functions of SFRP2. The Western blot, real-time RT-PCR and ChIP assays were used to evaluate the mechanism of SFRP2. RESULTS: Under inflammation and hypoxia conditions, the over-expression of SFRP2 could enhance the osteo/odontogenic differentiation ability. Mechanismly, SFRP2 inhibited canonical Wnt/ß-catenin signalling pathway and then inhibited the target genes of nuclear factor kappa B (NFkB) signalling pathway. Inflammation or hypoxia conditions could promote the expression of lysine demethylase 2A (KDM2A) and repress SFRP2 transcription through decreasing histone methylation in the SFRP2 promoter. Besides, proteomic analysis showed that SFRP2 promoted SCAPs to secret more functional cytokines, which improve the migration, chemotaxis and osteo/odontogenic ability of MSCs. CONCLUSIONS: Our discoveries revealed that SFRP2 enhanced the osteo/odontogenic differentiation and paracrine potentials of SCAPs under hypoxia and inflammation conditions and provided a potential cytokine for promoting tissue regeneration in hypoxia and inflammatory niche.

2.
Pestic Biochem Physiol ; 160: 30-39, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31519255

RESUMO

An exploration of novel control strategies for Leptinotarsa decemlineata is becoming more pressing given rapid evolution of insecticide resistance and rise of production loss of potato. Dietary delivery of bacterially expressed double-stranded RNA (dsRNA) is a promising alternative for management. An important first step is to uncover possible RNA-interference (RNAi)-target genes effective against both young and old larvae. Taiman (Tai) is a basic-helix-loop-helix/Per-Arnt-Sim transcription factor that is involved in the mediation of both juvenile hormone (JH) and 20-hydroxyecdysone (20E) signaling. In the present paper, we found that continuous ingestion of dsTai for three days by third (penultimate)-instar larvae caused approximately 20% larval mortality and 80% pupation failure. The larval lethality resulted from failed cuticle and tracheae shedding, which subsequently reduced foliage consumption and nutrient absorption, and depleted lipid stores. In contrast, pupation failure derived from disturbed JH and 20E signals, and disordered nutrient homeostasis including, among others, inhibition of trehalose metabolism and reduction of chitin content. Knockdown of LdTai caused similar larval lethality and pupation impairment in second and fourth (final) larval instars. Therefore, LdTai is among the most attractive candidate genes for RNAi to control L. decemlineata larvae.

3.
Artigo em Inglês | MEDLINE | ID: mdl-31525721

RESUMO

Isoelectric focusing (IEF) has been used for determination of meat quality with high stability analysis. However, it still suffered from time-consuming, laborious and cost-effective performances, e.g., 3 h protein extraction, more than 10 h rehydration time, 5-12 h focusing time, and imaging of protein band. To overcome these issues, a speedy extraction of colorful proteins was developed by controlling extraction and centrifugation of 0.2g sample within 10 min and 15 min respectively; a rapid analytical method was designed by using a quick array IEF with 25 min rehydration, 7 min focusing, 2 min online scanning and imaging of focused proteins. The total analytical time was well controlled within 1 h, significantly less than the traditional IEF time of 24 h. To demonstrate the proposed method, 18 chickens were classified into three groups, e.g., the normal slaughtering, death treatment underwater, and death with infection via the New castle disease (NDV) virus. The experiments demonstrated that two Mb bands with pI 6.8 and 7.4 were present in slaughtered chickens, while four other bands with pI 6.83, 6.95, 7.09, and 7.13 were observed in abnormal chicken. The additional four proteins bands were identified by western blot (WB) as hemoglobin proteins. Furthermore, array Immobilized pH Gradient (IPG) has high sensitivity (absolute LOD of Mb and Hb were 1.3 ng and 5.5 ng), fair stability (RSD values of 2.32%, 2.27%, and 1.69%) for slaughtered, drowned, NDV-infected chickens for intra-day and (2.94%, 1.66%, and 1.07%) for inter-days, and good recovery (100%, 98.25% and 99.75%). Finally, the developed method could be used for the identification of chicken meat quality with less time and small volume reagents consuming.

4.
Hum Genet ; 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31535215

RESUMO

Premature ovarian insufficiency (POI) is a major cause of reduced female fertility and affects approximately 1% women under 40 years of age. Recent advances emphasize the genetic heterogeneity of POI. Fanconi anemia (FA) genes, traditionally known for their essential roles in DNA repair and cytogenetic instability, have been demonstrated to be involved in meiosis and germ cell development. Here, we conducted whole-exome sequencing (WES) in 50 Han Chinese female patients with POI. Rare missense variants were identified in FANCA (Fanconi anemia complementation group A): c.1772G > A (p.R591Q) and c.3887A > G (p.E1296G). Both variants are heterozygous in the patients and very rare in the human population. In vitro functional studies further demonstrated that these two missense variants of FANCA exhibited reduced protein expression levels compared with the wild type, suggesting the partial loss of function. Moreover, mono-ubiquitination levels of FANCD2 upon mitomycin C stimulation were significantly reduced in cells overexpressing FANCA variants. Furthermore, a loss-of-function mutation of Fanca was generated in C57BL/6 mice for in vivo functional assay. Consistently, heterozygous mutated female mice (Fanca+/-) showed reduced fertility and declined numbers of follicles with aging when compared with the wild-type female mice. Collectively, our results suggest that heterozygous pathogenic variants in FANCA are implicated in non-syndromic POI in Han Chinese women, provide new insights into the molecular mechanisms of POI and highlight the contribution of FANCA variants in female subfertility.

5.
Appl Opt ; 58(19): 5301-5309, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31503629

RESUMO

The optical behavior of twisted nematic liquid crystals (TNLCs) is revealed through an angular scanning technique. Experimental results show that the optical rotation and degree of polarization of transmitted light are dependent on the polarization direction of incident light. The optical rotation is reciprocal, i.e., the polarization direction of incident and transmitted light can reciprocate when optical rotation is π/2. In some cases, the optical rotation is zero. The orientation of alignment layers in the TN cell can be determined from the behavior of optical rotation, which agrees with the measurement by an atomic force microscope. The experimental results are explained with the model of circularly polarized light based on the circular birefringence effect. Linearly polarized incident light is the superposition of right- and left-handed circularly polarized light. The propagation velocity of circularly polarized light in the LC is relevant to the polarization direction of incident light, so that the refractive indices of left- and right-handed circularly polarized light, n- and n+, or circular birefringence Δn(=n--n+) are not constants. As a result, when a linearly polarized light with the wavelength λ propagates through a TN cell with the cell gap l, the polarization direction of transmitted light is rotated to an angle Δθ. The optical rotation Δθ(=π(n--n+)l/λ) is dependent on the polarization direction of incident light, whereas the averaged refractive index ⟨n⟩(=(n-+n+)/2) can be independent of that. The incident light is partially linearly polarized light in our experiments, so that the degree of polarization of transmitted light varies with the polarization direction of incident light because the optical rotatory rates for the primary and secondary light beams are different.

6.
ACS Nano ; 2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31483608

RESUMO

Excitons in semiconductors are usually noninteracting and behave like an ideal gas, but may condense to a strongly correlated liquid-like state, i.e., electron-hole liquid (EHL), at high density and appropriate temperature. An EHL is a macroscopic quantum state with exotic properties and represents the ultimate attainable charge excitation density in steady states. It bears great promise for a variety of fields such as ultra-high-power photonics and quantum science and technology. However, the condensation of gas-like excitons to an EHL has often been restricted to cryogenic temperatures, which significantly limits the prospect of EHLs for use in practical applications. Herein we demonstrate the formation of an EHL at room temperature in monolayer MoS2 by taking advantage of the monolayer's extraordinarily strong exciton binding energy. This work demonstrates the potential for the liquid-like state of charge excitations to be a useful platform for the studies of macroscopic quantum phenomena and the development of optoelectronic devices.

7.
Artigo em Inglês | MEDLINE | ID: mdl-31493080

RESUMO

The Southern Junggar Basin (SJB) in China is an emerging coalbed methane (CBM) development area with abundant low-rank CBM resources. CBM development is accomplished by pumping significant volumes of water from the aquifer, and this water is commonly termed as produced water, which has great utilization values for the water-deficient areas like SJB. Geochemistry signatures are prerequisites in the management of the produced water. Meanwhile, geochemistry surveys of this produced water could also help study the basin hydrogeology and then serve the CBM development. In this study, geochemical compositions of the produced waters, including major ions, stable isotopic compositions, trace elements, and rare earth elements, were analyzed. Results show that produced waters from CBM wells in the SJB are of Na-HCO3 type and have wide total dissolved solid (TDS) ranges from 963 to 11,916 mg/L (avg. 7417 mg/L). Cl-, Na+, and HCO3- are the principal determinates of the TDS contents of the produced waters, and their concentrations all increase with greater depth of the produced waters. Overall, the net results of groundwater-aquifer mineral-bacteria interactions with groundwater flowing along the flow path are to deplete Ca2+, Mg2+, and SO42- and increase Na+, Cl-, HCO3-, and TDS. Stable isotopic values of the CBM produced waters (δDH2O and δ18OH2O) cluster along or below the local meteoric water line (LMWL), and the shift of stable isotopic values to the right side of LMWL was affected by a joint effect of evaporation and mixing with near-surface water. Trace elements that exceed the regulated concentrations for drinking water of China include As, Fe, Mn, Ba, and Ni, among which Ba and Fe need to be most concerned because over 50% of the CBM produced waters exceed the regulated values. Through principal component analysis, the trace element associations in the CBM produced waters and their potential origins were analyzed. The ∑REY concentrations of the CBM produced waters increase exponentially with the increase of pH and present a certain correlation with TDS. The relationship between ∑REY concentrations and TDS reflects different water-rock reaction degrees and hydrogeological backgrounds.

8.
Org Lett ; 21(19): 7938-7942, 2019 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-31553199

RESUMO

We have successfully developed a green and efficient multicomponent reaction protocol to synthesize S-aryl dithiocarbamates under visible light. Most appealingly, the reaction can proceed smoothly without adding any transition-metal catalysts, ligands, or photocatalysts while minimizing chemical wastes and metal residues in the end products. The advantages of this method meet the requirements of sustainable and green synthetic chemistry, and it provides a straightforward way to create valuable S-aryl dithiocarbamates.

9.
Biomed Pharmacother ; 118: 109313, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31545233

RESUMO

Bone marrow-derived mesenchymal stem cells (MSCs) from systemic lupus erythematosus patients (SLE-BMSCs) exhibited abnormalities in cytokine production and immune modulation. Deregulation of Nod-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome plays an important role in SLE. Herein, we explored whether miRNAs are involved in the regulation of NLRP3 in SLE-BMSCs. ELISA assay was used to detect the levels of inflammatory cytokines. The expression levels of let-7f-5p and gene mRNAs were determined by qRT-PCR assay. The protein levels of NLRP3, Cleaved caspase-1 and ASC were measured by western blot. The interaction between let-7f-5p and NLRP3 was verified using dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. In vivo assay was performed to explore whether let-7f-5p upregulation could ameliorate inflammation in MRL/lpr mice. Our data indicated that SLE patients had significantly serum higher levels of IFN-γ, IL-6, IL-18, IL-12, IL-13 and IL-1ß. We demonstrated that NLRP3 expression was upregulated in SLE-BMSCs. Let-7f-5p directly targeted NLRP3 and repressed NLRP3 expression. NLRP3 depletion or let-7f-5p upregulation repressed IL-1ß production and the expression of NLRP3 inflammasome components. Moreover, upregulated let-7f-5p-mediated anti-inflammation effect was significantly abrogated by NLRP3 expression restoration. Besides, let-7f-5p upregulation ameliorated inflammation through modulating NLRP3 in vivo. In conclusion, our study suggested that high level of let-7f-5p alleviated inflammation in SLE-BMSCs at least partly through targeting NLRP3, highlighting let-7f-5p as a novel promising therapeutic strategy for SLE treatment.

10.
Parasitol Int ; 73: 101974, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31421266

RESUMO

Hookworms are blood-sucking nematodes that infect dogs, cats, and humans, causing iron-deficiency anemia, abdominal pain, diarrhea, and skin inflammation. Amplification refractory mutation system (ARMS) is a modified technology based on allele-specific PCR, which is widely used in mutation detection and genotyping. However, no data about ARMS application in hookworm detection. This study aims to establish a multi-ARMS-qPCR method for the detection of three hookworm species from dogs and cats. A universal forward primer and three specific primers (ARMS-Cey, ARMS-Can, and ARMS-Tub) were designed based on the three ITS SNPs (ITS250, ITS78 and ITS153) of Ancylostoma ceylanicum, A. caninum, and A. tubaeforme, respectively. The results showed that the three designed ARMS primers generated specific melting curves for the three hookworms' standard plasmids. The melting temperature (Tm) values were 88.40 °C (A. ceylanicum), 83.15 °C (A. caninum), and 85.65 °C (A. tubaeforme), with good reproducibility of intra- and inter-assay. No amplification was observed with other intestinal parasites. The limit of detection using the established technique was 1, 2, and 104 egg per gram feces (EPG) for A. caninum, A. tubaeforme and A. ceylanicum, respectively. Using multi-ARMS-qPCR assay, 17 out of 50 fecal samples were positive for hookworms, including ten single and seven mixed infections, and single infections were quantified. In conclusion, the used multi-ARMS-qPCR method has the advantages of high efficiency, sensitivity, specificity, and quantitative analysis and can be used for the clinical detection, epidemiological investigation, and zoonotic risk assessment of canine and feline hookworms.

11.
Int J Mol Sci ; 20(16)2019 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-31426343

RESUMO

Four kinds of Ca-montmorillonite with different layer charge density were used to study the effect of charge density on their hydration properties by molecular dynamics simulation and experiments. The research results of Z-density distribution of water molecules, Hw (hydrogen in water molecules), and Ca in the interlayer of montmorillonite show that the hydration properties of montmorillonite are closely related to its layer charge density. If the charge density is low, the water molecules in the interlayers are mainly concentrated on the sides of the central axis about -1.3 Å and 1.5 Å. As the charge density increases from 0.38semi-cell to 0.69semi-cell, the water molecules are distributed -2.5 Å and 2.4 Å away from the siloxane surface (Si-O), the concentration of water molecules near the central axis decreases, and at the same time, Ca2+ appears to gradually shift from the vicinity of the central axis to the Si-O surface on both sides in the montmorillonite layer. The simulation results of the radial distribution function (RDF) of the Ca-Hw, Ca-Ow (oxygen in water molecules), and Ca-Ot (the oxygen in the tetrahedron) show that the Ca2+ and Ow are more tightly packed together than that of Hw; with the increase of the charge density, due to the fact that the negative charge sites on the Si-O surface increase, under the action of electrostatic attraction, some of the Ca2+ are pulled towards the Si-O surface, which is more obvious when the layer charge density of the montmorillonite is higher. The results of the RDF of the Ot-Hw show that with the increase of charge density, the number of hydrogen bonds formed by Ot and Hw in the interlayers increase, and under the action of hydrogen bonding force, the water molecules near the central axis are pulled towards the two sides of Si-O surface. As a result, the arrangement of water molecules is more compact, and the structure is obvious. Correspondingly, the self-diffusion coefficient shows that the higher the layer charge density, the lower the self-diffusion coefficient of water molecules in interlayers is and the worse the hydration performance of montmorillonite. The experimental results of the experiments fit well with the above simulation results.

12.
Carcinogenesis ; 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31430374

RESUMO

Rho GTPase-activating proteins (RhoGAPs) have been reported to be of great importance in the initiation and development of many different cancers. However, their biological roles and regulatory mechanisms in lung cancer development and progression are poorly defined. Real-time PCR or western blotting analysis was used to detect Rho GTPase-activating protein 24 (ARHGAP24), WWP2, P27, p-STAT6 and STAT6 expression levels as well as the activity of RhoA and Rac1 in lung cancer. Cell proliferation, apoptosis and cell cycle were respectively measured by CCK-8 and flow cytometry analysis. Tumor growth of lung cancer cells was measured using a nude mouse xenograft experiment model in vivo. The correlation between WWP2 and P27 was measured by Co-immunoprecipitation and ubiquitination analysis. We found that ARHGAP24 expression was lower in lung cancer tissues collected from the TCGA and independent hospital database, respectively. Overexpression of ARHGAP24 significantly suppressed cell proliferation and the activity of RhoA and Rac1, induced cell apoptosis and arrested cell cycle at the G0-G1 phase. ARHGAP24 overexpression also inhibited tumor growth in nude mice. While, knockdown of ARHGAP24 significantly promoted cell proliferation and WWP2 expression and inhibited cell cycle arrest at G1 phase through activating STAT6 signaling. ARHGAP24 overexpression inhibited WWP2 overexpression-induced cell proliferation, cell cycle progression and the decreased p27 expression. Moreover, WWP2 was found interacted with P27, and WWP2 overexpression promoted the ubiquitination of P27. In conclusion, our findings suggest that ARHGAP24 inhibits cell proliferation and cell cycle progression and induces cell apoptosis of lung cancer via a STAT6-WWP2-P27 axis.

13.
J Cell Mol Med ; 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31411001

RESUMO

Rheumatoid arthritis (RA) is a chronic inflammation mediated by autoimmune responses. MEG3, a kind of long noncoding RNA (lncRNA), participates in cell proliferation in cancer tissues. However, the correlation between MEG3 and RA is yet unclear. Therefore, to clarify how MEG3 works in RA, we performed a series of experiments using RA samples. We found that MEG3 was downregulated in the fibroblast-like synoviocytes of RA patients (RA-FLS), in comparison with healthy subjects. MEG3 was also down-regulated evidently in lipopolysaccharide (LPS)-treated chondrocyte. As part of our experiments, MEG3 was overexpressed in chondrocyte by transfection with lentivirus containing sequences encoding MEG3. In addition, in presence of LPS, reductions were identified not only in the cell proliferation, but also in the generation of interleukin-23 (IL-23), which, however were reversed in the lentivirus (containing MEG3-encoding sequences)-transfected chondrocytes. Up-regulated MEG3 resulted in an increase the level of Ki67. Moreover, MEG3 was negatively correlated with miR-141, and miR-141 was up-regulated in LPS-treated chondrocyte. Inhibitory effects of MEG3 overexpression, mentioned above, were partially abolished by overexpressed miR-141. Further, animal experiment also showed the inhibitory effect of MEG3 in overexpression on the AKT/mTOR signaling pathway. In-vivoexperiments also showed that cell proliferation was facilitated by MEG3 overexpression with inhibited inflammation. In summary, the protective role of MEG3 in RA was proved to be exerted by the increase in the rate of proliferation, which might correlate to the regulatory role of miR-141 and AKT/mTOR signal pathway, suggesting that MEG3 holds great promise as a therapeutic strategy for RA.

14.
J Oral Rehabil ; 2019 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-31469431

RESUMO

Mesenchymal stem cell-based therapy is a reliable treatment for periodontal tissue regeneration, while ideal regeneration rate is still a facing problem. In previous study, we found SFRP2 a promising gene in modulating mesenchymal stem cells potential. We further investigated its role on periodontal tissue regeneration. We created periodontitis model in miniature pigs and locally injected with stem cells from apical papilla (SCAP). The periodontitis models were classed into three groups, SFRP2-SCAP group (injected with SCAP overexpressing with SFRP2), SCAP group (injected with SCAP transduced with vector backbone) and saline group (vehicle group injected with saline). Clinical assignment, CT scanning, histopathological assessment and quantitative analysis were applied to evaluate the regeneration effect. Twelve weeks after the injection, we found healthier gingival status in SFRP2-SCAP group than the other two groups. Clinical assignment results showed values of probing depth, gingival recession and attachment loss were improved in SFRP2-SCAP group than that of SCAP group and saline group. The volume of newborn bone was also enhanced in SFRP2-SCAP group than SCAP group and saline group. The difference of clinical assignments and newborn bone between each group was significant relevant. HE staining demonstrated increased tissue regeneration in SFRP2-SCAP group than SCAP group and saline group. Our findings revealed that SFRP2 could enhance SCAP-mediated periodontal tissue regeneration and provide a potential target for improving the regeneration of periodontal tissue.

15.
Vet Parasitol Reg Stud Reports ; 17: 100316, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31303229

RESUMO

Hookworm infection is globally prevalent among dogs and cats representing a major public health risk. Although previous studies have surveyed canine and feline hookworms in Guangzhou city, the status of these infection needs to be further explored in other regions of South China. To investigate the prevalence and zoonotic risk of canine and feline hookworms in eight cities (Guangzhou, Foshan, Shenzhen, Huizhou, Zhongshan, Shaoguan, Shantou and Chaozhou) of Guangdong province, China, we developed specific PCR methods based on ITS sequence for identifying three common hookworm species. The results showed that the prevalence of hookworms from stray dogs and cats was 20.23% (142/702) and 15.26% (47/308), respectively. The established PCR methods could identify Ancylostoma ceylanicum, A. caninum and A. tubaeforme. The mixed infections of A. caninum and A. ceylanicum were detected in stray dogs of Guangzhou and Shaoguan, with the rate of 8.3% and 21.2%, respectively. Among the stray dogs in Foshan, the infection rate of A. ceylanicum was higher than that of A. caninum. The stray cats in four of five investigated cities were infected with A. ceylanicum. The different region, age and rearing environments had an impact on the hookworm infection rates of stray dogs and cats. In conclusion, the reported higher infection rate of A. ceylanicum than other hookworm species in stray dogs and cats poses a potential risk to public health.

16.
Plant Dis ; 103(9): 2385-2391, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31313639

RESUMO

Stimulatory effects of low doses of fungicides on the virulence of phytopathogens have profound implications for applications of fungicides. The present study demonstrated that carbendazim sprayed at 0.001 to 0.03 µg/ml had stimulatory effects on the virulence of mycelia of Botrytis cinerea, and the maximum percent stimulations were 15.5 and 21.4% for isolates HB459 and HB536, respectively. Potato dextrose agar (PDA) amended with carbendazim at 0.01, 0.02, and 0.05 µg/ml inhibited mycelial growth of isolate HB536 by 0.8, 10.0, and 30.6%, respectively. However, after the inhibited mycelia were inoculated on cucumber leaves, virulence increased by 10.1, 12.9, and 10.8%, respectively. With respect to sclerotial production, carbendazim at 0.005 and 0.02 µg/ml in PDA significantly (P < 0.05) increased, while at 0.1 µg/ml significantly (P < 0.05) reduced the sclerotial number and weight of both isolates compared with nontreated controls. Conidia germination percentages slightly yet statistically significantly (P < 0.05) increased after being inoculated on PDA amended with carbendazim at 0.001 and 0.005 µg/ml. Carbendazim at 0.001∼0.02 µg/ml, either sprayed on cucumber leaves or cosuspended with conidia, exerted significantly (P < 0.05) stimulatory effects on the virulence of B. cinerea conidia. Mechanism studies showed that sublethal doses of carbendazim did not increase the expression levels of pathogenicity-related pectin methylesterase gene Bcpme1, endopolygalacturonase gene Bcpg2, cutinase gene CutA, xylanase gene Xyn11A, or NADPH oxidase gene BcnoxA.


Assuntos
Benzimidazóis , Botrytis , Carbamatos , Doenças das Plantas , Virulência , Benzimidazóis/farmacologia , Botrytis/efeitos dos fármacos , Carbamatos/farmacologia , Virulência/efeitos dos fármacos
17.
Electrophoresis ; 2019 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-31172555

RESUMO

A novel analytical protocol was developed for general quality screening of chicken meat based on IEF and protein extraction. To demonstrate the developed protocol, 24 chickens were divided into three groups; each had eight chickens. The chickens in Group 1 were slaughtered by exsanguination, Group 2 asphyxiated in water, and that in Group 3 were infected by new castle disease virus. Proteins were extracted from the meat samples by using pure water as an extractant, separated by IEF, verified by western blot, and quantified via imaging analysis. The relevant experiments demonstrated that two myoglobin (Mb) bands were detected at pI 6.8 and 7.04 for all samples of Groups 1, 2, and 3, but there were additional hemoglobin (Hb) bands at pI 7.09 and 7.13 (P < 0.05) for the samples of Groups 2 and 3. The results implied that Hb bands might be a potential biomarker for the screening of chicken meat quality. The RSD values of two Mb bands (pI 6.8 and 7.04) in Group 1 were respectively 4.08 and 3.63%, the ones of two Hb bands (pI 7.09 and 7.13) in Group 2 were 3.66 and 2.10%, and those in Group 3 were 2.17% and 2.77%, respectively. All the RSD values indicated high stability and reliability of the developed protocol. Additionally, the protocol had a direct readout of protein bands in IEF without staining. However, it was time-consuming and had high cost. Even so, the relevant general method and finding have potential for screening of chicken meat quality.

18.
J Cell Physiol ; 234(12): 22260-22271, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31081124

RESUMO

To better understand the molecular mechanisms of anaplastic thyroid carcinoma (ATC), we aimed to identify the hub genes specifically involved in ATC by integrated bioinformatics analysis. In this study, using three Gene Expression Omnibus data sets with the same platform GPL570, we screened hub genes involved in ATC progression. In vitro experiments, such as western blot analysis, Transwell assays, and coimmunoprecipitation, was performed to verify our findings. By comparing three subtypes of thyroid cancer with normal tissue, we found ATC harbored more changed genes than well and poorly differentiated thyroid cancer. Using specifically differentially expressed genes between ATC and normal thyroid tissues to perform Gene ontology (GO) analysis, ATC showed enrichments of GO terms involved in lymphocyte migration and activation, collagen catabolic and metabolic process, thyroid hormone synthesis, and embolism. Using genes involved in extracellular matrix, coexpression network analysis and protein-protein interaction analysis were performed to identify matrix metalloproteinase 3 (MMP3) and MMP13 as two hub genes. Our experimental data indicated that both MMP3 and MMP13 were upregulated in ATC and knockdown of either of them could notably suppress ATC cell invasion and migration. Mechanistically, Gene Set Enrichment Analysis, coimmunoprecipitation, and rescue experiments revealed MMP3 and MMP13 not only interacted with each other, but also regulated each other through the janus kinase/signal transducer and activator of transcription 3 and mammalian target of rapamycin pathways. In conclusion, we identified a specific molecular mechanisms for the development of ATC by integrated analysis of transcriptome and in vitro experiments, which suggested that MMP3 and MMP13 might be developed as novel therapeutic targets for ATC.

19.
Anal Chem ; 91(12): 7500-7504, 2019 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-31132248

RESUMO

Absolute quantification of ligand capped on the surface of nanoparticles (NPs) has faced a great challenge without the use of complex inner standards (CIS). Herein, we proposed a facile electrophoresis titration (ET) model, designed an ET device, and developed a relevant method for counting the ligand on NPs without the use of CIS, based on moving reaction boundary (MRB). Furthermore, we conducted the relevant ET runs by using 3-mercaptopropionic acid (MPA) and quantum dots (QDs) as the model ligand and NPs, respectively. The experiments revealed that the ligand content of 1518 ± 295 obtained via an ET was close to the one of 1408 ± 117 determined via NMR, validating the ET model. Moreover, the experiments showed fair stability (RSD < 5.62%) and simplicity of ET without the use of CIS. Evidently, the ET model opens a window for facile assay of ligand capped on NPs.

20.
Aging (Albany NY) ; 11(10): 3348-3361, 2019 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-31141496

RESUMO

We aimed to investigate the regulation of circular RNAs in lipopolysaccharide (LPS)-treated chondrocytes isolated from SD rat. In this study, we analyzed how circFADS2 was regulated in LPS-treated chondrocytes and isolates from Rheumatoid arthritis (RA) patients and found that circFADS2 and mTOR were highly expressed whereas miR-498 expression was significantly reduced. We then silenced circFADS2 in LPS-treated chondrocytes; this resulted in a declined expression of type II collagen, but an increase in the expression of MMP-13, COX-2, and IL-6. Overall, silencing circFADS2 caused a significant reduction in the proliferative rate of LPS-treated chondrocytes, increased apoptotic levels, miR-498 upregulation, and mTOR downregulation. Dual-luciferase reporter assay indicated that circFADS2 directly targeted miR-498. In contrast, miR-498 down-regulation affected circFADS2 silencing, promoting extracellular matrix (ECM) degradation and apoptosis. The 3' UTR of the mTOR gene is targeted by miR-498, and consequently, in cells transfected with miR-498, there was a significant reduction of mTOR expression at the protein and mRNA levels. Silencing mTOR had a similar effect to circFADS2 silencing on type II collagen, MMP-13, COX-2, and IL-6 expression, as well as cell proliferation and apoptosis. In conclusion, circFADS2 may affect LPS-induced chondrocytes properties by regulating the ECM catabolism, inflammation, and apoptosis in chondrocytes.

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