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1.
Int J Biol Macromol ; 2020 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-32224177

RESUMO

GIBP, a homogeneous polysaccharide extracted from Glycyrrhiza inflata Batalin with a molecular weight 1.96 × 103 kDa, had a triple helix structure, smooth and sheet-like structure. Comprehensive analysis showed that the main chain of GIBP was composed of α-D-1,4 linked glucose, branch points were composed of α-D-1,3,6 and α-D-1,2,3,6 linked glucoses, and side chains were composed of α-D-1,3 and ß-D-1,6 linked galactose, ß-L-1,2 linked arabinose, α-D-1,3 and ß-D-1,3 mannose. The scavenging abilities of GIBP (3 mg/mL) against DPPH radical, OH radical, O2- radical and ABTS were 50.75 ±â€¯0.13% and 52.32 ±â€¯0.13, 25.84 ±â€¯0.35% and 44.57 ±â€¯0.15% and it also demonstrated an obvious dose-effect relationship. The inhibitory activity of α-glucosidase showed that the inhibitory effect of GIBP was enhanced with the increase of concentration. When the concentration reached 6 mg/mL, the inhibition rate of α-glucosidase activity reached 64.77%. And the ka, kd and KD were 6.472 × 104 1/Ms., 2.934 × 10-3 1/s and 4.534 × 10-8 M.

2.
Zhen Ci Yan Jiu ; 45(1): 77-82, 2020 Jan 25.
Artigo em Chinês | MEDLINE | ID: mdl-32144914

RESUMO

The electrical characteristics of meridians and acupoints have been being one of the hot topics discussed by scholars, because of many influencing factors and inconsistent research results. In the present article, we collected papers published in Chinese or English in recent 10 years from PubMed, CNKI, and VIP databases by using key words of"acupuncture""meridian""resistance""capacitance""electrodermal"and"impedance". Then, we make an analysis about its development from three aspects, 1) manifestations of electrical properties of acupoints (low dermal resistance under pathological state, imbalance between left and right sides of the synonym acupoints), 2) clinical application of acupoint electrical characteristics for diagnosis of clinical disorders and evaluation of therapeutic effect, and 3) current situations of development of meridian detection instruments based on electrical characteristics (improving stability of instruments,expansion of the detected regions and measuring time, realization of visualization, and operational automation). In the process of coming studies, we highly recommend that more attention should be paid to the acupoints' "dynamic changes", and forming a standardized research plan, strengthening domestic and international academic exchanges and cooperation are definitely necessary in order to validate the objectivity and specificity of electrical characteristics of meridians and acupoints, further expanding its clinical application.


Assuntos
Pontos de Acupuntura , Meridianos , PubMed
3.
Zhen Ci Yan Jiu ; 45(2): 157-63, 2020 Feb 25.
Artigo em Chinês | MEDLINE | ID: mdl-32144928

RESUMO

OBJECTIVE: To investigate the specific regularity of body surface resistance at different acupoints of the meridians associated with the uterus in reflecting menstrual cycle by observing the change in body surface resistance at source points, cleft points, confluent points, and non-specific points of three yin meridians of the foot in different menstrual cycles in normal female college students, to lay a foundation for the in-depth research on the mechanism of acupoints reflecting the function of zang-fu, and to provide a reference for the clinical and scientific research on the biophysical characteristics of menstrual cycle-related acupoints in normal female. METHODS: A total of 90 normal female college students were recruited. The source points, cleftpoints, confluent points, and non-specific points of three yin meridians of the foot which were located in the adjacent spinal segments of the uterus were selected, and body surface resistance was monitored for 30 consecutive minutes at the same time-points of menstrual phase, follicular phase, ovulation phase, and luteal phase to observe the change in the resistance of each acupoint during the menstrual cycle. RESULTS: There was no significant change in the resistance value of unilateral acupoints during the menstrual cycle (P>0.05). In the same period, there was no significant difference in resistance value between unilateral three source points, three cleftpoints, different acupoints of the spleen meridian, and different acupoints of lumbar 4 dermatomere (P>0.05). As for the comparison of resistance of the same acupoint at the left and right sides, Taichong (LR3) at the left side had a higher resistance value than that at the right side in the menstrual phase (P<0.05); Taibai (SP3) at the left side had a higher resistance value than that at the right side in the ovulation phase (P<0.05); Zhongdu (LR6) at the left side had a higher resistance value than that at the right side in the follicular phase, the ovulation phase, and the luteal phase (P<0.05); Taixi (KI3), Diji (SP8), and Sanyinjiao (SP6) at the left side had a higher resistance value than those at the right side in the menstrual phase, the follicular phase, the ovulation phase, and the luteal phase (P<0.05); Shuiquan (KI5) at the left side had a lower resistance value than that at the right side in these four phases (P<0.05). CONCLUSION: The change trend of the resistance of the acupoints at the left and right sides associated with the three yin meridians of the foot can specifically reflect the change in qi and blood in the uterus during the menstrual cycle. The change trend of the source and cleft points of the liver meridian in the menstrual phase is different from that in the other phases, and the change trend of SP3, a source point of the spleen meridian, in the ovulation phase is different from that in the other phases, which suggests the specificity of meridian points in reflecting function. The mechanism by which meridian points reflect the function of zang-fu is associated with the meridian points and the spinal cord segments of zang-fu, and meanwhile, it has a specific relationship with the meridians to which meridian points belong and the attributes of acupoints.


Assuntos
Pontos de Acupuntura , Meridianos , Feminino , Humanos , Medula Espinal , Útero
4.
Carbohydr Polym ; 236: 116009, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32172837

RESUMO

In this study, whey protein isolate (WPI)-quercetin (Que)-lotus root amylopectin (LRA) hydrogels (WPI-QUE-LRA) was developed to improve the solubility, stability and bioavailability of quercetin. Results showed that the favorable WPI-QUE-LRA was formed using WPI and LRA in the ratio of 1:2 at pH 7.0. Under this condition, the average size, polydispersity index, zeta potential of the WPI-QUE-LRA was 179.5 nm, 0.271, -18.6 mV, respectively. The analysis of transmission electron microscopy, fourier transform infrared spectroscopy and X-ray diffractometer revealed that the quercetin was successfully encapsulated in WPI-LRA, giving a high encapsulation efficiency of 92.4 %. Moreover, the WPI-LRA could significantly improve the storage stability and photochemical stability of quercetin. The in vitro and in vivo experiments showed that LRA-coated WPI hydrogel can enable quercetin to be stable in stomach and be effectively released in small intestine, leading to the enhancement of the bioavailability of quercetin.

5.
Carbohydr Res ; 491: 107911, 2020 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-32217360

RESUMO

A novel GH36 α-galactosidase gene (LrAgal36A) from Lichtheimia ramosa was synthesized and highly expressed in Pichia pastoris. The enzyme titer and protein yield for high-density fermentation in a 5 L fermentor were up to 953.6 U mL-1 and 4.36 g L-1. Purified recombinant LrAgal36A showed the maximum activity at pH 6.0 and 65 °C and was thermostable with a half-life of 70 min at 60 °C. LrAgal36A displayed the highest specific activity (353.17 ± 4.19 U mg-1) toward p-nitrophenyl-α-d-galactopyranoside (pNPGal) followed by galacto-oligosaccharides and could act slightly on galactomannans. The Km and catalytic efficiency (kcat/Km) of LrAgal36A for pNPGal were 0.33 mM and 1569.50 mM-1 s-1, respectively. LrAgal36A and GH5 ß-mannanase from L. ramosa showed a significant synergistic effect on the degradation of locust bean gum (LBG), resulting in release more reducing sugars (1.56 folds) and galactose (7.6 folds) by simultaneous or sequential reactions. Due to its hydrolysis properties, LrAgal36A might have potential applications in the area of pulp biobleaching, feed and food processing.

6.
Allergy ; 2020 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-32198890

RESUMO

BACKGROUND: The contribution of B-cell subsets and T-B cell interaction to the pathogenesis of allergic rhinitis (AR) and mechanisms of allergen immunotherapy (AIT) remain poorly understood. This study aimed to outline circulating B-cell signature, the underlying mechanism, and its association with clinical response to AIT in patients with AR. METHODS: IgD/CD27 and CD24/CD38 core gating systems were used to determine frequencies and phenotypes of B cells. Correlations between B cells, T cells, antigen-specific IgE, and disease severity in AR patients were investigated. Switched memory B cells were co-cultured with type 2 follicular helper T (Tfh2) cells and follicular regulatory T (Tfr) cells. Associations between B-cell subsets and clinical benefits of AIT were analyzed. RESULTS: Frequencies and absolute numbers of circulating memory B cells were increased in AR patients. CD23 expression on CD19+ CD20+ CD27+ IgD- switched memory B cells was significantly enhanced and positively correlated with antigen-specific IgE levels, symptom scores, and Tfh2/Tfr cell ratio in AR patients. Compared to those from healthy controls, Tfh2 cells from AR patients had a greater capacity to induce CD23 expression on switched memory B cells via IL-4, which was unable to be sufficiently suppressed by AR-associated Tfr cells with defective IL-10 expression. CD23 expression on switched memory B cells was downregulated after 12-month AIT, which positively associated with disease remission in AR patients. CONCLUSION: T-B cell interaction, bridged by CD23 expression particularly on switched memory B cells, may be involved in the disease pathogenesis and mechanism of AIT in patients with AR.

8.
Zhen Ci Yan Jiu ; 45(3): 237-42, 2020 Mar 25.
Artigo em Chinês | MEDLINE | ID: mdl-32202717

RESUMO

OBJECTIVE: To investigate the disease spectrum and predominant diseases treated by abdominal acupuncture by data mining and analysis of journal articles on abdominal acupuncture, and to provide a reference for clinical diagnosis and treatment. METHODS: Based on the database of abdominal acupuncture established by the research group, the data mining technique was used for the analysis and extraction of the articles on abdominal acupuncture included in this study. RESULTS: A total of 788 original journal articles were included. Six departments and 96 disease categories, among which there were 45 internal diseases (46.84%) and 18 surgical diseases (18.75%) were involved. As for the diseases involved, cervical spondylosis had the highest frequency of 84, followed by low back and leg pain with a frequency of 77 and stroke sequela with a frequency of 67. Of all 788 studies, 519 (65.86%) used abdominal acupuncture combined with other therapies with a total frequency of 552, among which acupuncture had the highest frequency of 135 (24.46%), followed by oral administration of traditional Chinese medicine with a frequency of 81 (14.67%) and moxibustion with a frequency of 80 (14.49%). Abdominal acupuncture had a marked clinical effect in the treatment of various diseases, with the highest effective rate of 95.10% in surgical diseases. CONCLUSION: Abdominal acupuncture has a wide disease spectrum and is most frequently used for the treatment of cervical spondylosis, with a marked clinical effect. Abdominal acupuncture has unique therapeutic characteristics and advantages, but it can achieve a better clinical effect when combined with other therapies.

9.
Molecules ; 25(3)2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32019202

RESUMO

The importance of the gut microbiota in drug metabolism, especially in that of nonabsorbable drugs, has become known. The aim of this study was to explore the metabolites of triptolide by the gut microbiota. With high-performance liquid chromatography coupled with tandem mass spectrometry and ion trap time-of-flight multistage mass spectrometry (LC-MS/MS and LC/MSn-IT-TOF), four metabolites of triptolide (M1, M2, M3, and M4) were found in the intestinal contents of rats. M1 and M2, were isomeric monocarbonyl-hydroxyl-substituted metabolites with molecular weights of 390. M3 and M4 were isomeric dehydrogenated metabolites with molecular weights of 356. Among the four metabolites, the dehydrogenated metabolites (M3 and M4) were reported in the gut microbiota for the first time. The metabolic behaviors of triptolide in the gut microbiota and liver microsomes of rats were further compared. The monocarbonyl-hydroxyl-substituted metabolites (M1 and M2) were generated in both systems, and another monohydroxylated metabolite (M5) was found only in the liver microsomes. The combined results suggested that the metabolism of triptolide in the gut microbiota was specific, with two characteristic, dehydrogenated metabolites. This investigation might provide a theoretical basis for the elucidation of the metabolism mechanism of triptolide and guide its proper application in clinical administration.

10.
BMC Ophthalmol ; 20(1): 45, 2020 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-32013911

RESUMO

BACKGROUND: To investigate the prevalence of pterygium and associated factors in Han and Mongolian adults at four survey sites in Inner Mongolia, China. METHODS: We conducted a population-based, cross-sectional study as part of the China National Health Survey (CNHS). By means of a stratified sampling method, we finally included 2651 participants of 30 years of age or older from a total of 3468 eligible residents. Factors associated with pterygium were analysed by a univariate analysis and logistic regression models. RESULTS: The study population included 1910 Han and 741 Mongolian adults. The mean age ± standard deviation of the study cohort was 48.93 ± 11.06 years. The overall prevalence of pterygium was 6.4% (n = 169); 1.4% (n = 38) of the cases were bilateral and 4.8% (n = 128) were unilateral. The most common grade of pterygium was Grade 2. Based on the results of the univariate analysis, eleven factors were included in a multivariate analysis. The results indicated that age (P < 0.001), outdoor occupation (P = 0.026), and time spent in rural areas (P < 0.001) were significantly associated with pterygium. Sex and ethnicity were not identified as risk factors. CONCLUSIONS: Our results indicated that outdoor occupation, old age and more time spent in rural areas were risk factors for pterygium in Inner Mongolia. At the same time, town as a survey site (Hohhot and Tsining District) was a protective factor for pterygium. Ethnicity, gender, smoking, diabetes and high blood pressure are not associated with pterygium.

11.
Biomed Res Int ; 2020: 9204708, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32047820

RESUMO

Chronic hypoxia (CH) is characterized by long-term hypoxia that is associated with microvessel proliferation and basal membrane (BM) degradation in tissues. The IL-6/JAK2/STAT3/MMP-9 pathway has been described in a variety of human cancers and plays an essential role in microvessel proliferation and BM degradation. Therefore, this study investigated the role of the IL-6/JAK2/STAT3/MMP-9 pathway in hypoxia-mediated microvessel proliferation and BM degradation in the rat bone marrow. Eighty pathogen-free Sprague Dawley male rats were randomly divided into four groups (20 per group)-control group, CH group (exposed to hypoxia in a hypobaric chamber at a simulated altitude of 5000 m for 28 d), CH + STAT3 inhibitor group (7.5 mg/kg/d), and CH + DMSO group. Microvessel density (MVD) and BM degradation in the bone marrow were determined by immunofluorescence staining and transmission electron microscopy. Serum IL-6 levels were assessed by enzyme-linked immunosorbent assay (ELISA), and the levels of P-JAK2, P-STAT3, and MMP-9 were assessed by western blot analysis and real-time reverse transcription PCR (RT-PCR). Hypoxia increased serum IL-6 levels, which in turn increased JAK2 and STAT3 phosphorylation, which subsequently upregulated MMP-9. Overexpression of MMP-9 significantly promoted the elevation of MVD and BM degradation. Inhibition of STAT3 using an inhibitor, SH-4-54, significantly downregulated MMP-9 expression and decreased MVD and BM degradation. Surprisingly, STAT3 inhibition also decreased serum IL-6 levels and JAK2 phosphorylation. Our results suggest that the IL-6/JAK2/STAT3/MMP-9 pathway might be related to CH-induced microvessel proliferation and BM degradation in the bone marrow.

12.
Microb Pathog ; 142: 104061, 2020 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-32061916

RESUMO

To compare different nanoparticle-based nasal vaccines against foot-and-mouth disease (FMD), chitosan (CS)-coated poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) (CS/PLGA-NPs) and amino-functionalized mesoporous silica nanoparticles (Am/MSNs) loaded with FMDV recombinant plasmid (pP12A3C/IFN-CS/PLGA-NPs and pP12A3C/IFN-Am/MS-NPs) were used to induce mucosal and systemic immune responses in guinea pigs via intranasal delivery. Simultaneously, CpG oligodeoxy nucleotides (ODNs) as a vaccine adjuvant were encapsulated in chitosan-coated poly (lactic-co-glycolic acid) nanoparticles (CpG-CS/PLGA-NPs). The pP12A3C/IFN-CS/PLGA-NPs and CpG-CS/PLGA-NPs generated displayed good morphology, high stability, mean diameters of 500 and 400 nm and encapsulation efficiencies of 83.8% and 88.4%, respectively. Data from the in vitro release assay showed that plasmid and CpG were sustainably released from nanoparticles (up to 66.73% and 64%, respectively, of the total amount loaded). Guinea pigs immunized with pP12A3C/IFN-CS/PLGA-NPs + CpG-CS/PLGA-NPs showed markedly higher mucosal, cellular and humoral immune responses than those administered pP12A3C/IFN-CS/PLGA-NPs or naked plasmid vaccine alone. FMDV-specific secretory immunoglobulin A (sIgA) antibodies in nasal washes were initially detected at 3 days post-vaccination with CS/PLGA-NPs loaded with plasmid. Guinea pigs immunized with pP12A3C/IFN-CS/PLGA-NPs also displayed higher cellular and humoral immune responses than pP12A3C-CS/PLGA-NPs and naked plasmid vaccine alone. FMDV-specific immunoglobulin G (IgG) antibodies in serum were initially detected at 5 days post-vaccination (intramuscularly) with the naked plasmid. Finally, challenge experiments 42 days post-vaccine revealed 100% protection in guinea pigs immunized with pP12A3C/IFN-CS/PLGA-NPs + CpG-CS/PLGA-NPs and pP12A3C/IFN-CS/PLGA-NPs. However, plasmid DNA was burst released from pP12A3C/IFN-Am/MS-NPs. Our attempts to use pP12A3C/IFN-Am/MS-NPs to immunize guinea pigs failed to induce immune responses. In conclusion, CpG and IFN-α adjuvant based FMD vaccines elicit protection in guinea pigs. Moreover, CS-coated PLGA NPs present an efficient and safe mucosal immune delivery system for FMDV DNA vaccine. Data from the current study provide a foundation for understanding and further evaluating protective immune responses in pigs.

13.
J Agric Food Chem ; 68(10): 3112-3120, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32046486

RESUMO

Maternal gut dysbiosis affects the development of the offspring immune system. Our previous study has indicated that microbial metabolite butyrate directly shapes pancreatic immune tolerance and dampens type 1 diabetes (T1D) progression. Therefore, maternal butyrate intervention may protect their offspring from maternal gut dysbiosis-accelerated T1D. To test this, pregnant nonobese diabetic (NOD) mice were treated with vancomycin in drinking water with or without a butyrate-supplemented diet during gestation and nursing (oral vancomycin is used to induce maternal gut dysbiosis). Three weeks after delivery, T1D-associated innate and adaptive immune cells were detected to investigate the effects of butyrate on the vancomycin-exacerbated pancreatic immune disorder in dams and pups. The results showed that butyrate inhibited maternal vancomycin-exacerbated secretion of proinflammation cytokines (interferon γ and interleukin-1ß) and maternal vancomycin-exacerbated recruitment of interferon γ+ T cells (cytotoxic T lymphocytes 1 cells and T helper type 1 cells) in the pancreas of the female offspring, thus dampening T1D development. The protection may be due to butyrate inhibiting the activation of pancreatic dendritic cells (DCs). Our data thus demonstrate that maternal gut dysbiosis can exacerbate pancreatic-directed autoimmunity in the female offspring through T cell- and DC-associated mechanisms that are inhibited by butyrate.


Assuntos
Antibacterianos/efeitos adversos , Butiratos/administração & dosagem , Diabetes Mellitus Tipo 1/tratamento farmacológico , Vancomicina/efeitos adversos , Animais , Células Dendríticas/imunologia , Diabetes Mellitus Tipo 1/induzido quimicamente , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/imunologia , Feminino , Humanos , Interferon gama/genética , Interferon gama/imunologia , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Masculino , Camundongos , Camundongos Endogâmicos NOD , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia
14.
J Vis Exp ; (156)2020 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-32090987

RESUMO

Begomoviruses (genus Begomovirus, family Geminiviridae) are transmitted by whiteflies of the Bemisia tabaci complex in a persistent, circulative manner. Considering the extensive damage caused by begomoviruses to crop production worldwide, it is imperative to understand the interaction between begomoviruses and their whitefly vector. To do so, localization and quantification of the virus in the vector tissues is crucial. Here, using tomato yellow leaf curl virus (TYLCV) as an example, we describe a detailed protocol to localize begomoviruses in whitefly midguts, primary salivary glands, and ovaries by immunofluorescence. The method is based on the use of specific antibodies against a virus coat protein, dye-labeled secondary antibodies, and a confocal microscope. The protocol can also be used to colocalize begomoviral and whitefly proteins. We further describe a protocol for the quantification of TYLCV in whitefly midguts, primary salivary glands, hemolymph, and ovaries by quantitative PCR (qPCR). Using primers specifically designed for TYLCV, the protocols for quantification allow the comparison of the amount of TYLCV in different tissues of the whitefly. The described protocol is potentially useful for the quantification of begomoviruses in the body of a whitefly and a virus-infected plant. These protocols can be used to analyze the circulation pathway of begomoviruses in the whitefly or as a complement to other methods to study whitefly-begomovirus interactions.

15.
J Proteomics ; : 103691, 2020 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-32068187

RESUMO

Acute myocardial infarction (AMI) is an acute heart disease. Cycloastragenol, as a natural product, inhibits inflammation and protects cardiomyocytes. Cycloastragenol (Y006) modulates inflammation in AMI is not known. To explore the function of Cycloastragenol in AMI, this study investigated the effect of Y006 and its mechanisms both in vitro and in vivo. Y006 influences the concentration of 11 proteins, as shown by a proteomics analysis, immunohistochemistry and western blotting. Among these 11 proteins, Erk1/2, PLCG1, IKBKG, and ZEB1 are related to inflammatory regulation. BAX, COX2, and GSK3ß are involved in modulating cardiomyocyte apoptosis, and RhoA and DSC2 are directly associated with myocardial function. However, the functions of ARHGAP17 and Rit2 in heart are less well established. Additionally, Y006 suppressed TNF-α, IFN-γ and IL-17 production in PBMCs (peripheral blood monocytes) from patients with acute myocardial infarction and enhanced IL-10 and IL-4 expression. Similar results were obtained in a rat model of AMI by flow cytometry detection and ELISA. Our findings indicate that Y006 protects rats from AMI through direct or indirect inhibition of inflammation and cardiomyocyte apoptosis. However, the specific mechanism of Y006's protective function requires further study. Nonetheless, this research revealed a novel aspect for the treatment of myocardial infarction. SIGNIFICANCE: In the present study, we undertook the first proteomic evaluation of Cycloastragenol (Y006) function in acute myocardial infarction (AMI). Y006 significantly improved myocardial function in vivo by regulating multiple molecular expressions. Hypoxia is a direct reason for AMI. And our data support a role of Y006 in gene expression, cell apoptosis under hypoxia. The conclusions of this research assist to explain the potential molecular mechanism in Cycloastragenol treating AMI and supply a new method for ameliorating AMI.

16.
Int J Biol Macromol ; 148: 591-600, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31958563

RESUMO

The present work aims to investigate the effects and underlying mechanism of a homogeneous Laminaria japonica polysaccharide (LJP61A) on acute kidney injury (AKI) in mice. According to the results of biochemical and pathological analysis, we concluded that LJP61A could protect kidney from the damage of adriamycin in AKI mice. Compared to the model group, the mRNA level of cytokines (TNF-α, IL-1ß and MCP-1) and protein level of mesenchymal markers demsin were decrease by the treatment of LJP61A while the protein levels of podocyte structure markers (Nephrin and WT-1) were increased. Moreover, the adriamycin-induced enhancement of phosphor-p65, phosphor-p38, phosphor-ERK1/2 and phosphor-JNK in the kidney of AKI mice were significantly suppressed by LJP61A. Similar variation was observed in the mRNA and protein levels of TGF-ß1 and Smad3. These results suggested that LJP61A prevented acute kidney injury possibly via regulating TGF-ß1-mediated Smad3, MAPKs and NF-κB signaling pathways.

17.
Chemistry ; 2020 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-31953882

RESUMO

MicroRNAs (miRNAs) modulate the expression of over 30 % of mammalian genes during development and apoptosis, and abnormal expression of miRNAs may lead to a range of human pathologies. Therefore, analysis of miRNAs is valuable for disease diagnostics. In this work, a novel one-pot fluorescence derivatization strategy was developed for miRNA analysis. The mechanism of the derivatization reaction was explored by using instrumental methods, including liquid chromatography, fluorescence spectroscopy, and mass spectrometry. Highly fluorescent N6 -ethenoadenine (ϵ-adenine) was formed and detached from the miRNA sequence through the reaction of adenine in nucleic acids with 2-chloroacetaldehyde (CAA) at 100 °C. This is the first experimental evidence that the cooperation of formed ϵ-adenine and water-mediated hydrogen-bond interaction between the proton at the 2'- and the oxyanion at 3'-positions stabilized the oxocarbenium significantly, which makes the depurination and derivatization of miRNA highly effective. Based on this derivatization strategy, a facile and sensitive high-performance liquid chromatography method was developed for quantitative assay of miRNAs. In combination with magnetic solid-phase extraction (MSPE), the HPLC method was shown to be useful for the determination of microRNAs at sub-picomolar level in serum samples.

18.
Biotechnol Lett ; 42(4): 613-624, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31970554

RESUMO

OBJECTIVE: To survey genome-scale protease profiles regulated by the Aspergillus niger transcription factor PrtT and further controlled by carbon sources. RESULTS: The PrtT disruption mutant (delprtT) and overexpression (OEprtT) strains were successfully generated and further confirmed by phenotypic and protease activity analysis. RNA-seq analysis of WT and mutants identified 32 differentially expressed protease genes, which mostly belonged to serine-type peptidases, aspartic-type endopeptidases, aminopeptidases and carboxypeptidases. Furthermore, based on the MEME predicted motif analysis of the PrtT promoter, EMSA and phenotypic and qRT-PCR analyses confirmed that the carbon metabolism regulator AmyR directly regulated the protease genes and their regulatory factor PrtT. CONCLUSION: Thirty-two PrtT-regulated protease genes were identified by RNA-seq, and the secondary carbon source regulator AmyR was found to have a negative regulatory effect on the expression of PrtT and its target protease genes.

19.
Br J Pharmacol ; 2020 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-31976546

RESUMO

BACKGROUND AND PURPOSE: Despite recent advances in understanding its pathophysiology, treatment of acute kidney injury (AKI) remains a major unmet medical need, and novel therapeutic strategies are needed. Cathelicidin-related antimicrobial peptide (CRAMP) with immunomodulatory properties has an emerging role in various disease contexts. Here, we aimed to investigate the role of CRAMP and its underlying mechanisms in AKI. EXPERIMENTAL APPROACH: The human homologue LL-37 and CRAMP were measured in blood samples of AKI patients and in experimental AKI mice respectively. Experimental AKI was induced in wild-type and CRAMP-deficient (Cnlp-/- ) mice by ischaemia/reperfusion (I/R). Therapeutic evaluation of CRAMP was performed with exogenous CRAMP (5 mg·kg-1 , i.p.) treatment. KEY RESULTS: Cathelicidin expression was inversely related to clinical signs in patients and down-regulated in renal I/R-induced injury in mice. Cnlp-/- mice exhibited exacerbated I/R-induced renal dysfunction, aggravated inflammatory responses and apoptosis. Moreover, over-activation of the NLRP3 inflammasome in Cnlp-/- mice was associated with I/R-induced renal injury. Exogenous CRAMP treatment markedly attenuated I/R-induced renal dysfunction, inflammatory response and apoptosis, correlated with modulation of immune cell infiltration and phenotype. Consistent with Cnlp-/- mouse data, CRAMP administration suppressed renal I/R-induced NLRP3 inflammasome activation, and its renal protective effects were mimicked by a specific NLRP3 inhibitor CY-09. The reno-protective and NLRP3 inhibitory effects of CRAMP required the EGF receptor. CONCLUSION AND IMPLICATIONS: Our results suggest that CRAMP acts as a novel immunomodulatory mediator of AKI and modulation of CRAMP may represent a potential therapeutic strategy.

20.
Clin Exp Rheumatol ; 2020 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-31969221

RESUMO

OBJECTIVES: Takayasu's arteritis (TAK) involves inflammatory vasculitis of large vessels and mainly affects the aorta and its major branches. Abnormal immunity may play a vital role in TAK pathogenesis. Regulatory T cells (Treg cells) are important for peripheral tolerance, but under certain conditions Treg cells can differentiate into Th-like cells that have lost immune suppressive function and promote the development of autoimmune diseases. The role of Th-like Treg cells in TAK is unclear and this study aims to investigate the function of Th-like Treg cell subsets and associated cytokines in TAK. METHODS: A total of 51 patients with TAK and 32 healthy controls were enrolled. The percentage of Th1, Th2, Th17, Tregs and Th-like Treg cells in blood samples was analyzed by flow cytometry. Serum cytokine levels were detected using a cytometric bead array for cytokines. RESULTS: TAK patients had decreased numbers of Th2-like Treg cells in the peripheral blood (p=0.002) relative to healthy controls. The percentage of Treg cells in samples from TAK patients also decreased (p=0.002), but the Th2 cell percentage (p=0.04) increased compared to healthy controls. TAK patients had higher serum levels of IL-4 (p<0.001) and IL-13 (p<0.001) than healthy controls, and levels of both cytokines correlated to IL-6 levels. CONCLUSIONS: We studied changes in T helper-like Treg cell subsets in TAK for the first time and discovered that the number of Th2-like Treg cells in peripheral blood decreased. Results of this study suggested that Th2-like Treg cells could contribute to TAK pathogenesis.

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