Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 152
Filtrar
1.
Interv Neuroradiol ; : 1591019919897765, 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31969074

RESUMO

OBJECTIVE: Subclavian steal syndrome results from hemodynamic impairment due to stenosis or occlusion of subclavian artery. Therefore, it is important for subclavian steal syndrome patients to assess hemodynamic status during endovascular therapy. METHODS: Eleven subclavian steal syndrome patients undergoing endovascular therapy attended this study. Pressure wire was used to measure trans-stenosis pressure difference (ΔP). Digital subtraction angiography, Transcranial Doppler and Electronic sphygmomanometer were introduced to assess stenotic rate, steal grade and inter-arm systolic pressure difference, respectively. Clinical symptoms and restenosis were followed up after endovascular therapy. The associations of ΔP with stenotic rate, inter-arm pressure difference, steal degree, clinic symptoms and restenosis were analyzed in this paper. RESULTS: Prior to the therapy, ΔP moderately correlated with stenotic rate (r = 0.757, p = 0.007) and inter-arm pressure difference (r = 0.701, p = 0.016). ΔP was ≥6 mmHg in all patients, and 6-9 mmHg for grade 1 steal and ≥10 mmHg for grade 2 and 3 steals. After the therapy, all patients had technique success, and 10 patients had clinic success, and 1 patient appeared restenosis. ΔP was ≤3 mmHg and steal disappeared in the patients with clinical success. ΔP was 18 mmHg and grade 3 steal still existed in one patient without clinical success. One patient with 1 mmHg of ΔP after therapy appeared restenosis in the follow-up. CONCLUSION: The trans-stenosis pressure difference is closely related to steal degree and clinical symptoms. The measurement of hemodynamic status by pressure wire is very useful to guide endovascular therapy in subclavian steal syndrome patients. However, the restenosis may still occur, even though the hemodynamic impairment is improved.

2.
J Asian Nat Prod Res ; : 1-10, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31791147

RESUMO

Various bioactive polyketides have been found in Aloe barbadensis. However, the polyketide synthases (PKSs), which participate in biosynthesis of polyketides in A. barbadensis remain unknown. In this study, two type III PKSs (AbPKS1 and AbPKS2) were identified from A. barbadensis. AbPKS1 and AbPKS2 were able to utilize malonyl-CoA to yield heptaketides (TW93a and aloesone) and octaketides (SEK4 and SEK4b), respectively. AbPKS1 also exhibited catalytic promiscuity in recognizing CoA thioesters of aromatics to produce unusual polyketides. What Is more, a whole cell biocatalysis system with the capability of producing 26.4 mg/L of SEK4/SEK4b and 2.1 mg/L of aloesone was successfully established.

4.
J Asian Nat Prod Res ; : 1-7, 2019 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-31738087

RESUMO

Bistachybotrysin K (1), one new phenylspirodrimane dimer with a central 6/7 oxygen heterocycle core, was isolated from the fungus Stachybotrys chartarum CGMCC 3.5365. Its structure was elucidated by extensive spectroscopic data and single-crystal X-ray diffraction. Compound 1 showed significant cytotoxicity against human tumor cell lines HCT116, NCI-H460, BGC823, Daoy, and HepG2 with IC50 values in the range of 1.1-4.7 µM.

5.
Medicine (Baltimore) ; 98(22): e15863, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31145339

RESUMO

RATIONALE: A Mason type III radial head fracture, which is characterized by comminuted fragments of the radial head, is a severe injury. Open reduction and internal fixation (ORIF) is an alternative treatment method; however, the technique of using an on-table reduction in combination with surgical glue is rarely reported. PATIENT CONCERNS: A 48-year-old man was admitted to our department with complaints of elbow pain after falling down. Elbow radiography and computed tomography (CT) demonstrated characteristics of fractures before the operation. DIAGNOSIS: Radiographic images showed a Mason type III radial head fracture. INTERVENTIONS: The patient underwent ORIF at our hospital. During the operation, the technique of on-table reconstruction combined with surgical glue was used. OUTCOMES: The patient recovered well and was able to participate in his usual work. LESSONS: Mason type III radial head fractures could be treated with ORIF, and a satisfactory result could be anticipated, thus avoiding a radial head replacement or resection. Anatomical reduction of a comminuted radial head could be obtained via an on-table reconstruction and application of surgical glue.


Assuntos
Cimentos para Ossos , Fraturas Cominutivas/cirurgia , Redução Aberta/métodos , Fraturas do Rádio/cirurgia , Fraturas Cominutivas/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Rádio (Anatomia)/patologia , Rádio (Anatomia)/cirurgia , Fraturas do Rádio/patologia , Resultado do Tratamento
6.
J Orthop Surg (Hong Kong) ; 27(2): 2309499019842879, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30987529

RESUMO

PURPOSE: The distal tibiofibular syndesmosis is an important structure for ankle stability. The objective of this study was to evaluate the motion of the syndesmosis under different loading patterns and determine the characteristics of the syndesmotic motion. METHODS: Six fresh cadaveric lower extremity specimens with the knee reserved were tested in this study. The skin and muscles were removed with all ligaments around the syndesmosis and knee and ankle joint intact. An axial load of 600 N was applied to the specimens with the ankle joint in 10° dorsiflexion, neutral position, and 15° plantar flexion using a universal material testing machine. Then, with the ankle joint positioned neutrally, a combination of 600-N axial and 5-Nm torsional external rotation loading was applied to the specimens. The medial-lateral and anterior-posterior displacement and rotation of the distal fibula relative to the distal tibia were measured. RESULTS: Under the axial loading, the distal fibula tended to move medially and anteriorly and rotate internally with the ankle positioned from the neutral position to 15° plantar flexion. Meanwhile, when the ankle was positioned from the neutral position to 10° dorsiflexion, the distal fibula tended to move laterally and posteriorly and rotate externally. Under the combined loading, with respect to the isolated axial loading, the distal fibula tended to move medially and posteriorly, and rotate externally relative to the distal tibia. CONCLUSION: Micro motion existed in the syndesmosis. The relative motion of the syndesmosis was correlated to the ankle position and loading patterns.

7.
Artigo em Inglês | MEDLINE | ID: mdl-30650034

RESUMO

Morpholine is a common chemical used as emulsifier in the preparation of wax coatings for some fruit to help them remain fresh and protect against insects and fungal contamination. It has been reported that morpholine has acute toxic effects on rodents. In the present study, morpholine concentrations were analysed in fruits (citrus fruits, apples, strawberries and grapes) and juices (apple juice and orange juice) in order to determine dietary exposure among the Chinese population. A total of 732 fruit and juice samples were collected during 2015-2016, which covered major foods in China. Fruit and juice consumption data were taken from China National Nutrient and Health Survey (2002) and include data from 16,407 fruit or juice consumers. It was found that mean dietary exposure to morpholine residues from fruits and/or juices for general Chinese consumers and children 2-6 years old were 0.42 and 1.24 µg/kg bw/day, respectively. The 97.5% intake in general Chinese consumers and children 2-6 years old were 2.25 and 6.90 µg/kg bw/day, respectively. The primary food sources of the morpholine dietary intake of general Chinese consumers were citrus fruits (57.4%) and apples (40.8%). These findings suggested that dietary exposure to morpholine in the Chinese population was lower than the acceptable daily intake of morpholine, and there are no health concerns.


Assuntos
Dieta/estatística & dados numéricos , Exposição Dietética , Contaminação de Alimentos/análise , Sucos de Frutas e Vegetais/análise , Frutas/química , Morfolinas/análise , China
8.
J Asian Nat Prod Res ; 21(9): 887-894, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30614271

RESUMO

Three new phenylspirodrimanes derivatives named stachybotrysins H and I (1 and 2) and stachybotrin E (3), together with one known compound stachybotrylactam (4), were isolated from Stachybotrys chartarum CGMCC 3.5365. Their structures were determined by extensive NMR data and mass spectroscopic analysis. Compounds 1 and 2 showed inhibitory effect towards potassium channel Kv1.3 with IC50 values of 13.4 and 10.9 µM, respectively.


Assuntos
Canal de Potássio Kv1.3/antagonistas & inibidores , Compostos de Espiro/química , Stachybotrys/química , Animais , Células CHO , Linhagem Celular , Cricetinae , Cricetulus
9.
Biomed Environ Sci ; 32(11): 823-838, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31910940

RESUMO

OBJECTIVE: To identify the important risk factors for type 2 Diabetes Mellitus (T2DM) and develop effective strategies to address the problem of T2DM. Our study aimed to evaluate the association between apolipoprotein E (ApoE) genetic polymorphism and type 2 diabetes, and to provide clues for the etiology of T2DM. METHODS: Based on the criteria of inclusion and exclusion, we extracted, pooled, analyzed and assessed the case-control studies of ApoE polymorphism and T2DM published in PubMed, Web of Science, Medline, WanFang, VIP, and CNKI databases by R soft-ware (version 3.4.3). We used Random-effect models when heterogeneity was present in between-study, and fixed-effect models otherwise. RESULTS: We had 59 studies covering 6,872 cases with T2DM and 8,250 controls, and compared the alleles and genotypes of ApoE between cases and controls. When we conducted a comparison between ApoE ε4 and ε3 alleles, we produced a pooled OR of 1.18 (95% CI: 1.09-1.28; P < 0.001). ApoE ε2/ε2 genotype displayed a possible association with T2DM (OR = 1.46; 95% CI: 1.11-1.93; P = 0.007), ε3/ε4 genotype showed a 1.11-fold risk (OR = 1.11; 95% CI: 1.01-1.22; P = 0.039) and ε4/ε4 genotype had a 1.71-fold risk of developing T2DM (OR = 1.71; 95% CI: 1.33-2.19; P < 0.001) when they were compared with ε3/ε3 genotype. CONCLUSION: There is an association between ApoE polymorphism and T2DM: allele ε4 and genotypes (ε2/ε2, ε3/ε4, and ε4/ε4) are associated with the increased risk for the development of T2DM, and they may be risk factors for T2DM.

10.
Med Sci Monit ; 24: 5398-5404, 2018 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-30074982

RESUMO

BACKGROUND The aim of this study was to evaluate the effects of carotid artery angioplasty and carotid artery stenting (CAS) on cerebral blood flow (CBF) and cerebrovascular reactivity (CVR) in patients with preoperative cerebrovascular hemodynamic impairment. MATERIAL AND METHODS Seventeen patients with unilateral severe internal carotid artery (ICA) stenosis and ipsilateral CVR impairment underwent CAS. CBF and CVR were measured by single photon emission computed tomography (SPECT) with inhalation of carbon dioxide (CO2) one week before and three months after CAS. Sixty-eight ROIs in the middle cerebral artery (MCA) territory were analyzed in 17 patients. RESULTS Before CAS, CVR was impaired in all ROIs. CBF was impaired in 16 ROIs (23.5%). The percentage of ROIs with impaired CBF was significantly increased in patients with ≥90% carotid artery stenosis (p=0.047) without collateral flow through the circle of Willis (p=0.005). CAS significantly increased CVR in ROIs with a normal preoperative CBF and impaired CVR, indicating mild hemodynamic impairment (0.9±6.7% vs. 4.9±8.6%) (p=0.014). CAS significantly increased CBF in ROIs with preoperative impaired CBF and impaired CVR, indicating severe hemodynamic impairment (79.1±7.5% vs. 86.7±10.0%) (p<0.001). Following CAS, ROIs with normal CBF and impaired CVR had a significantly increased percentage of improved CVR (p=0.047); ROIs with impaired CBF and impaired CVR had a significantly increased percentage of improved CBF (p=0.027). CONCLUSIONS The severity of preoperative hemodynamic impairment, which is related to the degree of carotid artery stenosis and cerebral collateral flow, may influence hemodynamic benefits by CAS.


Assuntos
Dióxido de Carbono/farmacologia , Estenose das Carótidas/diagnóstico por imagem , Tomografia Computadorizada de Emissão de Fóton Único/métodos , Idoso , Angioplastia/métodos , Encéfalo/irrigação sanguínea , Dióxido de Carbono/metabolismo , Artérias Carótidas/diagnóstico por imagem , Estenose das Carótidas/cirurgia , Circulação Cerebrovascular/fisiologia , China , Feminino , Hemodinâmica/fisiologia , Humanos , Masculino , Pessoa de Meia-Idade , Artéria Cerebral Média/diagnóstico por imagem , Stents
11.
Int J Chron Obstruct Pulmon Dis ; 13: 2183-2190, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30140150

RESUMO

Purpose: Previous studies have described the incidence, risk factors, and outcomes for patients with acute exacerbations of COPD (AECOPD) developing acute kidney injury (AKI). However, little is known about the differences between community-acquired AKI (CA-AKI) and hospital-acquired AKI (HA-AKI) in patients with AECOPD. Thus, in this study, we compared prevalence, risk factors, and outcomes for these patients with CA-AKI and HA-AKI. Patients and methods: This study was conducted from January 2014 to January 2017, and data from adult inpatients with AECOPD were analyzed retrospectively. A total of 1,768 patients were included, 280 patients were identified with CA-AKI and 97 patients were with HA-AKI. Results: Prevalence of CA-AKI was 15.8% and that of HA-AKI was 5.5%, giving an overall AKI prevalence of 21.3%. Patients with CA-AKI had a higher prevalence of chronic kidney disease (CKD) and lower prevalence of chronic cor pulmonale than patients with HA-AKI. Risk factors for developing HA-AKI and CA-AKI were similar, such as being elderly, requirement for mechanical ventilation, and a history of coronary artery disease and CKD. Patients with HA-AKI were more likely to have stage 3 AKI and worse short-term outcomes. In comparison with patients with CA-AKI, those with HA-AKI were more likely to require non-invasive mechanical ventilation (31.3% versus 16.8%; P = 0.003) and had a longer duration of mechanical ventilation (11 days versus 8 days; P = 0.020), longer hospitalization (14 days versus 12 days; P = 0.038), and higher inpatient mortality (32.0% versus 13.2%; P < 0.001). Patients with HA-AKI had worse (multivariate-adjusted) inpatient survival than those with CA-AKI (hazard ratio, 1.7 [95% confidence interval, 1.03-2.81; P = 0.038] for the HA-AKI group). Conclusion: AKI was common in patients with AECOPD requiring hospitalization. CA-AKI was more common than HA-AKI but otherwise demonstrated similar demographics and risk factors. Nevertheless, patients with HA-AKI had worse short-term outcomes.


Assuntos
Lesão Renal Aguda/etiologia , Progressão da Doença , Hospitalização , Doença Pulmonar Obstrutiva Crônica/complicações , Lesão Renal Aguda/epidemiologia , Lesão Renal Aguda/patologia , Idoso , Idoso de 80 Anos ou mais , China/epidemiologia , Feminino , Mortalidade Hospitalar , Humanos , Masculino , Prevalência , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Estudos Retrospectivos , Fatores de Risco
12.
J Asian Nat Prod Res ; 20(7): 605-614, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29989425

RESUMO

Glucuronidation is an important and popular metabolic reaction in vivo of drugs. The further evaluation of biological activity and toxicity of glucuronides is necessary in the course of the drug research and development. However, the synthesis of glucuronides is limited by the lack of efficient approach. Herein, we have developed a new glucuronide synthesis method using plant uridine diphosphate-dependent glucuronosyltransferases (UGTs), UGT88D4, UGT88D7, and EpGT8, enabling the convenient preparation for corresponding O-glucuronide metabolites (1a, 2a, 3a, and 3b) in milligram scale of two neurological active agents, IMM-H004 (1) and FLZ (2). Their structures were characterized by spectroscopic data analyses.


Assuntos
Glucuronídeos/síntese química , Glucuronosiltransferase/metabolismo , Plantas/enzimologia , Clonagem Molecular , Glucuronosiltransferase/química , Glucuronosiltransferase/genética , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância Magnética , Metais/química , Estrutura Molecular , Espectrometria de Massas por Ionização por Electrospray , Temperatura Ambiente
14.
J Asian Nat Prod Res ; 20(9): 844-851, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29119831

RESUMO

Two new lanostane triterpenoids (1 and 2), two new ergostane-type steroids (3 and 4) together with two known lanostane triterpenoids (5 and 6) and one known steroid (7) were isolated from the cultured mycelia of Ganoderma capense (CGMCC 5.71). Their structures were determined on the basis of extensive spectroscopic (HRESIMS, 1D NMR, 2D NMR) data analyses. Compound 1 exhibited moderate cytotoxic activity against the human cancer cell line NCI-H1650 with an IC50 value of 22.3 µM, and 7 displayed cytotoxic activity against the human cancer cell line HCT116 with an IC50 value of 17.4 µM. In addition, compounds 2, 3, 5, and 6 displayed weak anti-HIV activity with IC50 values of 23.5, 46.7, 21.6, and 30.1 µM, respectively.


Assuntos
Ganoderma/química , Micélio/química , Esteroides/química , Triterpenos/química , Ganoderma/metabolismo , Estrutura Molecular , Micélio/metabolismo , Esteroides/metabolismo
15.
Xenobiotica ; 48(3): 250-257, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28285550

RESUMO

1. UDP-glucuronosyltransferases (UGTs) are important drug-metabolizing enzymes (DMEs) catalyzing the glucuronidation elimination of various xenobiotics and endogenous substances. Endogenous substances are important regulators for the activity of various UGT isoforms. Triiodothyronine (T3) and thyroxine (T4) are important thyroid hormones essential for normal cellular differentiation and growth. The present study aims to elucidate the inhibition behavior of T3 and T4 on the activity of UGT isoforms. 2. In vitro recombinant UGTs-catalyzed glucuronidation of 4-methylumbelliferone (4-MU) was used to screen the inhibition potential of T3 and T4 on the activity of various UGT isoforms. Initial screening results showed that T4 exerted stronger inhibition potential than T3 on the activity of various UGT isoforms at 100 µM. Inhibition kinetics was determined for the inhibition of T4 on the representative UGT isoforms, including UGT1A1, -1A3, -1A7, -1A8, -1A10 and -2B7. The results showed that T4 competitively inhibited the activity of UGT1A1, -1A3, -1A7, 1A10 and -2B7, and noncompetitively inhibited the activity of UGT1A8. The inhibition kinetic parameters were calculated to be 1.5, 2.4, 11, 9.6, 4.8 and 3.0 µM for UGT1A1, -1A3, -1A7, -1A8, -1A10 and -2B7, respectively. In silico docking method was employed to demonstrate why T4 exerted stronger inhibition than T3 towards UGT1A1. Stronger hydrogen bonds and hydrophobic interaction between T4 and activity cavity of UGT1A1 than T3 contributed to stronger inhibition of T4 towards UGT1A1. 3. In conclusion, more clinical monitoring should be given for the patients with the elevation of T4 level due to stronger inhibition of UGT isoforms-catalyzed metabolism of drugs or endogenous substances by T4.


Assuntos
Glucuronosiltransferase/antagonistas & inibidores , Tiroxina/farmacologia , Tri-Iodotironina/farmacologia , Inibidores Enzimáticos/farmacologia , Glucuronosiltransferase/química , Glucuronosiltransferase/metabolismo , Humanos , Ligações de Hidrogênio , Himecromona/metabolismo , Simulação de Acoplamento Molecular , Tiroxina/química , Tri-Iodotironina/química
16.
Sci Rep ; 7(1): 17399, 2017 12 12.
Artigo em Inglês | MEDLINE | ID: mdl-29234143

RESUMO

Sleep apnoea is associated with chronic kidney diseases. A high obstructive sleep apnoea (OSA) prevalence is shown in patients with hypertrophic cardiomyopathy (HCM). Whether the presence of OSA would affect the renal function of patients with HCM is unknown. Forty-five consecutive patients with HCM were divided into the HCM OSA- and OSA+ groups. Forty-three patients with OSA without HCM were recruited as controls. Clinical indices, including estimated glomerular filtration rate (eGFR) and urine 8-hydroxy-2-deoxyguanosine (8-OHdG), were measured. The eGFR was significantly lower in the HCM OSA+ group than in the HCM OSA- (P < 0.05) and OSA (P < 0.001) groups. Multivariate linear regression analysis identified that the apnoea-hypopnoea index was independently associated with eGFR in all patients with HCM (ß = -1.329, 95% confidence interval: -1.942, -0.717, P < 0.001). The urine 8-OHdG level, an oxidative stress marker, was significantly higher in the HCM OSA+ group than in the HCM OSA- (P < 0.001) and OSA (P < 0.001) groups and significantly correlated with the AHI (r = 0.467, P = 0.003) and eGFR (r = -0.457, P = 0.004) in all patients with HCM. Our study suggests a risk of eGFR decline in patients with HCM and OSA.


Assuntos
Cardiomiopatia Hipertrófica/complicações , Insuficiência Renal Crônica/etiologia , Apneia Obstrutiva do Sono/complicações , Adulto , Idoso , Feminino , Taxa de Filtração Glomerular , Humanos , Masculino , Pessoa de Meia-Idade , Insuficiência Renal Crônica/epidemiologia , Fatores de Risco
17.
Am J Hypertens ; 30(12): 1203-1210, 2017 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-28992284

RESUMO

BACKGROUND: Homocysteine-lowering intervention with folate was recently shown to be able to increase day-night difference of blood pressure (BP) in humans indicating a potential relationship between homocysteine and circadian BP variation. We thus sought to investigate the association between plasma total homocysteine level (tHcy) and circadian BP variation in hypertensive adults. METHODS: We enrolled 244 eligible dipping and 249 nondipping BP status adults from 560 adults who were randomly sampled from 5,233 Chinese hypertensive adults who received ambulatory BP monitoring (ABPM). We further enrolled 390 adults with CC/CT genotypes of the methylenetetrahydrofolate reductase (MTHFR) and 79 TT genotype who received ABPM at the same time from 1858 hypertensive adults with MTHFR polymorphisms detection. RESULTS: Plasma tHcy in nondippers was significantly higher than dippers (P < 0.001). Simple linear analysis revealed that tHcy significantly correlated with nocturnal systolic BP fall (r = -0.145, P = 0.001) and diastolic BP fall (r = -0.141, P = 0.002). Multivariate logistic regression analysis further identified tHcy as an independent factor correlated with the presence of nondipping BP status in hypertensive adults (odds ratio: 1.873, 95% confidence interval: 1.171-2.996, P = 0.009). The percentage of dipping BP status was 19.49% or 8.86% and the percentage of nondipping BP status was 80.51% or 91.14% in CC/CT or TT genotypes, respectively. The above different between CC/CT and TT genotypes was significant (P = 0.024). CONCLUSIONS: These results indicated that high homocysteine levels associate with disturbed circadian BP variation in Chinese hypertensive adults.


Assuntos
Pressão Sanguínea , Ritmo Circadiano , Homocisteína/sangue , Hipertensão/sangue , Adulto , Idoso , Grupo com Ancestrais do Continente Asiático , Monitorização Ambulatorial da Pressão Arterial , Feminino , Genótipo , Humanos , Hipertensão/genética , Hipertensão/fisiopatologia , Masculino , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Pessoa de Meia-Idade , Polimorfismo Genético
18.
J Microbiol Biotechnol ; 27(11): 1971-1982, 2017 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-28910860

RESUMO

Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti-SpaA-based CIB procedure could quantitatively detect 2.5 × 103 CFU/ml of pLR colonies spiked in 106 CFU/ml of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.


Assuntos
Fezes/microbiologia , Proteínas de Fímbrias/imunologia , Microbioma Gastrointestinal/fisiologia , Soros Imunes/imunologia , Immunoblotting/métodos , Lactobacillus rhamnosus/imunologia , Lactobacillus rhamnosus/isolamento & purificação , Probióticos , Ácidos , Animais , Aderência Bacteriana/efeitos dos fármacos , Aderência Bacteriana/imunologia , Ácidos e Sais Biliares/farmacologia , Western Blotting , Células CACO-2 , Análise por Conglomerados , Reações Cruzadas/imunologia , Microbioma Gastrointestinal/efeitos dos fármacos , Microbioma Gastrointestinal/imunologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Lactobacillus rhamnosus/efeitos dos fármacos , Lactobacillus rhamnosus/genética , Proteínas de Membrana/química , Proteínas de Membrana/imunologia , Camundongos , Subunidades Proteicas , RNA Ribossômico 16S/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Proteínas Recombinantes/imunologia
19.
Life Sci ; 188: 186-191, 2017 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-28768154

RESUMO

Glycyrrhetinic acid (GA) is a natural active component from licorice, which is broadly used in traditional Chinese medicine. Lots of glycyrrhetinic acid receptors (GA-R) are proved to locate on the surface of liver cells. Many reports about the hepatocellular carcinoma (HCC) treatment were dependent on GA modified carriers. However, the reality of GA-R in HCC cells was not clear. In this paper, 18ß-glycyrrhetinic acid (18ß-GA) was labeled with fluorescence (FITC) by chemical synthesis. Together with the binding effect of fluorescence labeled glycyrrhetinic acid (FITC-GA), the competitive action of 18ß-GA with GA-R was investigated in HCC cells. The results showed that in HepG2 cells, 18ß-GA and FITC-GA presented similar cytotoxicity. The specific binding saturation of GA showed the dissociation constant (Kd) was 7.457±2.122pmol/L and the maximum binding counts (Bmax) was 2.385±0.175pmol/2.5×106 cells, respectively. FITC-GA bound to cytomembrane specifically and 18ß-GA competed to bind the sites significantly in HepG2 cells. Therefore, there is binding effect between fluorescence labeled GA and GA-R. The GA-R on HCC cells is confirmed as expected, which provides a useful reference of active target modified by GA and a novel approach for receptors and ligands study.


Assuntos
Carcinoma Hepatocelular/metabolismo , Ácido Glicirretínico/análogos & derivados , Ligantes , Apoptose/efeitos dos fármacos , Ligação Competitiva , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Fluoresceína-5-Isotiocianato/química , Fluoresceína-5-Isotiocianato/metabolismo , Fluoresceína-5-Isotiocianato/farmacologia , Ácido Glicirretínico/química , Ácido Glicirretínico/metabolismo , Ácido Glicirretínico/farmacologia , Humanos
20.
Zhongguo Zhong Yao Za Zhi ; 42(12): 2323-2328, 2017 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-28822187

RESUMO

The glycosides of 4'-demethylepipodophyllotoxin (DMEP) possess various pharmacological activities; however, the chemical synthesis of these glycosides faces challenges in regioselectivity, stereoselectivity, and the protection and de-protection of functional groups. In this work, a novel glycosyltransferase (GT) gene AbGT5 from Aloe barbadensis was successfully cloned, heterogeneously expressed and purified. Recombinant AbGT5 was able to catalyze the glycosylation of DMEP and the glycosylated product, which was separated from the preparative scale reaction, was characterized as DMEP 4'-O-ß-D-glucoside via MS, 1H-NMR, 13C-NMR, HSQC and HMBC. According to the investigations of enzyme properties, AbGT5 show the highest activity around 20 ℃ in the buffer of pH 9.0, and it was independent of divalent metal ions. Under the optimum conditions, the conversion rate of DMEP can reach 80%. Above all, in this work the enzymatic glycosylation of DMEP was achieved with high efficiency by the novel GT AbGT5.


Assuntos
Glucosídeos/química , Glicosídeos/química , Glicosiltransferases/metabolismo , Podofilotoxina/análogos & derivados , Aloe/enzimologia , Aloe/genética , Glicosilação , Glicosiltransferases/genética , Podofilotoxina/química
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA