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Various dyes are used to visualize DNA bands in agarose gel electrophoresis (AGE) by the methods of pre- or post-staining. The DNA dye user's guides generally state that the binding of the dye to DNA will affect DNA mobility in electrophoresis, thus recommending post-staining for accurate measurement of DNA size. However, many AGE performers prefer pre-staining procedures for reasons such as convenience, real-time observation of DNA bands, and/or the use of a minimal amount of dye. The detrimental effect of the dye on DNA mobility and the associated risk for inaccurate measurement of DNA size are often overlooked by AGE performers. Here we quantitatively determine the impact on DNA migration imposed by frequently used dyes, including GelRed, ethidium bromide (EB), and Gold View. It was observed that pre-staining with GelRed and EB significantly slowed down DNA migration to cause as much as 39.1% overestimation on the size of sample DNA, whereas Gold View had little effect. The slowdown of DNA migration increased with dye concentration until it plateaued when the dye concentration reached a saturated level. Thus, to take advantage of pre-staining, saturated levels of DNA dyes should always be applied for both DNA samples and DNA markers to ensure a fair comparison of DNA sizes. In addition, GelRed and EB display much higher sensitivity than Gold View in the detection of DNA bands in post-staining. The saturated concentrations, cost considerations, and other useful features of these frequently used dyes are summarized for the information of AGE performers.
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BACKGROUND: Venous complications after pediatric liver transplantation seriously affect the survival rate of patients and grafts. At present, the diagnostic indicators have not been unified. Venous complications may cause portal hypertension, which may lead to splenomegaly and splenic vein dilatation. Therefore, the changes in spleen may be closely related to the venous complications. The purpose of this study was to explore the relationship between ultrasonic splenic parameters and venous complications and to study whether these splenic parameters can be used for the diagnosis of venous complications. METHODS: We retrospectively included pediatric patients who underwent liver transplantation and collected ultrasonic spleen parameters before, and then 1-3 days, 1-3 weeks, 1-3 months, and 4-12 months after liver transplantation. We observed whether there were portal vein or hepatic vein complications within 1 year after liver transplantation. RESULTS: Among 109 pediatric patients after liver transplantation included in our study, 11 of them suffered from portal vein complications and nine hepatic vein complications. Spleen transverse diameter, spleen longitudinal diameter, spleen portal vein diameter, spleen index, spleen transverse diameter ratio, spleen longitudinal diameter ratio, and spleen index ratio were independent risk factors of venous complications. The accuracy of spleen transverse diameter (AUROC: 0.73), spleen index (AUROC: 0.70), spleen transverse diameter ratio (AUROC: 0.71), and spleen index ratio (AUROC: 0.72) in predicting venous complications were higher than other ones. CONCLUSIONS: Ultrasonic examination is a common follow-up method for pediatric patients after liver transplantation and the application of ultrasonic spleen parameters may be helpful to monitor venous complications.
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Transplante de Fígado , Baço , Humanos , Criança , Baço/diagnóstico por imagem , Transplante de Fígado/efeitos adversos , Transplante de Fígado/métodos , Estudos Retrospectivos , Veia Porta/diagnóstico por imagem , Ultrassonografia , Veia Esplênica/diagnóstico por imagemRESUMO
Au-Fe3O4 multifunctional nanoparticles (NPs) were synthesized and integrated with lateral flow immunoassay (LFIA) for dual-mode detection of Salmonella typhimurium. The Au-Fe3O4 NPs not only combined excellent local surface plasmon resonance characteristics and superparamagnetic properties, but also exhibited good photothermal effect. In the detection, antibody-conjugated Au-Fe3O4 NPs first captured S. typhimurium from complex matrix, which was then loaded on the LFIA strip and trapped by the T-line. By observing the color bands with the naked eyes, qualitative detection was performed free of instrument. By measuring the photothermal signal, quantification was achieved with a portable infrared thermal camera. The introduction of magnetic separation achieved the enrichment and purification of target bacteria, thus enhancing the detection sensitivity and reducing interference. This dual-mode LFIA achieved a visual detection limit of 5 × 105 CFU/mL and a photothermal detection limit of 5 × 104 CFU/mL. Compared with traditional Au-based LFIA, this dual-mode LFIA increased the detection sensitivity by 2 orders of magnitude and could be directly applied to unprocessed milk sample. Besides, this dual-mode LFIA showed good reproducibility and specificity. The intra-assay and inter-assay variation coefficients were 3.0% and 7.9%, and with this dual-mode LFIA, other bacteria hardly produced distinguishable signals. Thus, the Au-Fe3O4 NPs-based LFIA has potential to increase the efficiency of pandemic prevention and control. Au-Fe3O4 nanoparticle proved to be a promising alternative reporter for LFIA, achieving multifunctions: target purification, target enrichment, visual qualitation, and instrumental quantification, which improved the limitations of traditional LFIA.
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Nanopartículas Metálicas , Nanopartículas Multifuncionais , Salmonella typhimurium , Colorimetria , Reprodutibilidade dos Testes , ImunoensaioRESUMO
BACKGROUND: Histidine-tryptophan-ketoglutarate (HTK) and del Nido (DN) cardioplegia are intracellular-type and extracellular-type solution respectively, both can provide a long period of myocardial protection with single-dose infusion, but studies comparing the two are rare for adult cardiac surgery. This study aims to evaluate whether DN is suitable for cardioplegia in complex and high-risk valve surgery with long-term cardiac ischemia when compared with HTK. METHODS: The perioperative records of adult patients infused with DN/HTK as a cardioplegic solution who underwent complex valve surgery with an expected myocardial ischaemic duration longer than 90 min between Oct 2018 and Oct 2019 were analysed retrospectively. RESULTS: Of the 160 patients who received DN/HTK and underwent complex valve surgery, we propensity matched 73 pairs. Both groups achieved satisfactory cardiac arrest effects, and no significant difference was found in their cTnI and CK-MB levels within 12 to 72 h postoperatively. The DN group had a higher rate of return to spontaneous rhythm (0.88 v 0.52, P < 0.001), a lower frequency of postoperative severe arrythmias (12% v 26%, P = 0.036), a higher postoperative stroke volume (65 v 59 ml, P = 0.011) and a higher cardiac output (6.0 v 4.9 L/min, P = 0.007) as evaluated by echocardiography, fewer transfusions and shorter ICU stays (both P < 0.05). The two groups had similar inotrope usage and similar incidences of low cardiac output, morbidities and mortality. Subgroup analysis showed that when the aortic clamping time was greater than 120 min, the advantages of DN were weakened. CONCLUSIONS: DN can be safely applied to complex valve surgery, and it has a similar myocardial protection effect as HTK. Further prospective studies are required to verify these retrospective findings. Trial registration retrospectively registered.
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Procedimentos Cirúrgicos Cardíacos , Eletrólitos/administração & dosagem , Parada Cardíaca Induzida , Doenças das Valvas Cardíacas/cirurgia , Valvas Cardíacas/cirurgia , Lidocaína/administração & dosagem , Sulfato de Magnésio/administração & dosagem , Manitol/administração & dosagem , Cloreto de Potássio/administração & dosagem , Bicarbonato de Sódio/administração & dosagem , Soluções/administração & dosagem , Adolescente , Adulto , Idoso , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Eletrólitos/efeitos adversos , Feminino , Glucose/administração & dosagem , Glucose/efeitos adversos , Parada Cardíaca Induzida/efeitos adversos , Doenças das Valvas Cardíacas/diagnóstico por imagem , Doenças das Valvas Cardíacas/fisiopatologia , Valvas Cardíacas/diagnóstico por imagem , Valvas Cardíacas/fisiopatologia , Humanos , Lidocaína/efeitos adversos , Sulfato de Magnésio/efeitos adversos , Masculino , Manitol/efeitos adversos , Pessoa de Meia-Idade , Duração da Cirurgia , Complicações Pós-Operatórias/etiologia , Cloreto de Potássio/efeitos adversos , Procaína/administração & dosagem , Procaína/efeitos adversos , Recuperação de Função Fisiológica , Estudos Retrospectivos , Bicarbonato de Sódio/efeitos adversos , Soluções/efeitos adversos , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
Anthracnose fruit rot of litchi (Litchi chinensis Sonn.), caused by Colletotrichum spp., has been mainly associated with the C. acutatum species complex and C. gloeosporioides species complex (Farr and Rossman 2020). In June 2010, isolates of the C. acutatum species complex were isolated together with the C. gloeosporioides species complex from anthracnose lesions on litchi fruits (cv. Nuomici) obtained from a litchi orchard in Shenzhen (N 22.36°, E 113.58°), China. The symptoms typically appeared as brown lesions up to 25 mm in diameter, causing total fruit rot and sometimes fruit cracking. Based on the number of isolates we collected, the C. acutatum species complex appears less frequently on infected fruit compared to the C. gloeosporioides species complex. Since only the C. gloeosporioides species complex has been reported in China (Qi 2000; Ann et al. 2004), we focused on the C. acutatum species complex in this study. Pure cultures of fungal isolates were obtained by single-spore isolation. The isolate GBLZ10CO-001 was used for morphological characterization, molecular and phylogenetic analysis, and pathogenicity testing. Colonies were cultured on potato dextrose agar (PDA) at 25 â for 7 days, circular, raised, cottony, gray or pale orange, with reverse carmine, and 39.6 to 44.7 mm in diameter. Conidia were 13.5 to 19 × 4 to 6 µm (mean ± SD = 15.9 ± 1.1 × 5.2 ± 0.3 µm, n = 50) in size, hyaline, smooth-walled, aseptate, straight, fusiform to cylindrical with both ends acute. Appressoria were 5.5 to 13.5 × 4.5 to 7.5 µm (mean ± SD = 7.6 ± 1.6 × 6.0 ± 0.7 µm, n = 50) in size, subglobose to elliptical, sometimes clavate or irregular, smooth-walled, with entire edge, sometimes undulate, pale to medium brown. These morphological characteristics were consistent with the descriptions of several Colletotrichum species belonging to the C. acutatum species complex, including C. fioriniae (Shivas and Tan 2009; Damm et al. 2012). For molecular identification, genomic DNA was extracted and the ribosomal internal transcribed spacer (ITS), partial sequences of the ß-tubulin (TUB2), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase 1 (CHS-1), and histone3 (HIS3) genes were amplified and sequenced using the primer pairs ITS4/ITS5, T1/Bt2b, ACT512F/ACT783R, GDF1/GDR1, CHS-79F/CHS-354R, and CYLH3F/CYLH3R, respectively (White et al. 1990; Damm et al. 2012). The resulting sequences were submitted to GenBank (ITS: MN527186, TUB2: MT740310, ACT: MN532321, GAPDH: MN532427, CHS-1: MT740311, HIS3: MT740312). BLAST searches showed 98.70%-100% identity to the sequences of the C. fioriniae ex-holotype culture CBS 128517. The phylogram reconstructed from the combined dataset using MrBayes 3.2.6 (Ronquist et al. 2012) showed that isolate GBLZ10CO-001 clustered with C. fioriniae with high posterior probability. Koch's postulates were performed in the field to confirm pathogenicity. Isolate GBLZ10CO-001 was grown on PDA (25 â for 7 days) to produce conidia. In June 2014, litchi fruits (cv. Nuomici) were sprayed with conidial suspensions (106 conidia/ml), with sterile water as blank controls, and each treatment inoculated at least 15 fruits. Inoculated fruits were covered by an adhesive-bonded fabric bag until the trial ended. After 31 days, typical symptoms were observed, while control fruits remained asymptomatic. The fungus was re-isolated from diseased fruits and identified as C. fioriniae according to the methods described above. To our knowledge, this is the first report of anthracnose fruit rot on litchi caused by C. fioriniae, one species of the C. acutatum species complex, in China. For the difficulty in distinguishing anthracnose caused by C. fioriniae from the C. gloeosporioides species complex just by the symptoms, and mixed infection usually occurring in the field, further investigations are required to reliably assess the potential threat posed by C. fioriniae for litchi production in China.
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BACKGROUND: Processing of edible bird's nest (EBN) requires extensive washing to remove impurities and produces huge amounts of EBN co-products, which contain mainly feathers with glycoproteins attached, which are usually discarded. This study was conducted to recover the valuable EBN glycoproteins from the waste material. Enzymatic hydrolysis was applied to recover EBN glycopeptides from EBN co-products (EBNcoP ) and processed cleaned EBN (EBNclean ) was used as control, which were then freeze-dried into EBN hydrolysates (EBNhcoP and EBNhclean , respectively). RESULTS: The recovery yield for EBNhclean and EBNhcoP were 89.09 ± 0.01% and 47.64 ± 0.26%, respectively, indicating nearly 50% of glycopeptide can be recovered from the waste material. Meanwhile, N-acetylneuraminic acid, a major acid sugar in EBN glycoproteins, of EBNhcoP increased by 229% from 58.6 ± 3.9 to 192.9 ± 3.1 g kg-1 , indicating the enzymatic hydrolysis removed impurities and thus enhanced the N-acetylneuraminic acid content. Total soluble protein was more than 330 g kg-1 for all the samples. Colour parameter showed that hydrolysate samples have greater L* (lightness) values. Chroma result indicates the intensity of all the samples were low (< 11). Fourier-transform infrared (FTIR) spectrum displayed that the EBNhcoP exhibited similar functional groups with EBNhclean , indicating that the EBNcoP has similar functionality as EBNclean . Significantly higher (P ≤ 0.05) 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) activities were reported in EBNhcoP after the enzymatic reaction. CONCLUSION: EBNhcoP were successfully recovered from low value EBNcoP with enhanced antioxidant activities. The findings of this work are beneficial for the EBN industry to reduce wastage and enhance economic values of EBN co-products, both economically and nutritionally. © 2020 Society of Chemical Industry.
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Produtos Biológicos/química , Manipulação de Alimentos/métodos , Glicopeptídeos/química , Saliva/química , Animais , Antioxidantes/química , Antioxidantes/isolamento & purificação , Biocatálise , Produtos Biológicos/isolamento & purificação , Aves , Enzimas/química , Plumas/química , Glicopeptídeos/isolamento & purificação , HidróliseRESUMO
A new sorbent was synthesized for restricted-access matrix solid phase extraction (RAM-SPE) of the diabetes drugs metformin (MET) and glimepiride (Glim). Mesoporous silica layers were placed on Fe3O4-magnetized graphene modified with sulfo-functionalized pore walls (denoted as magG@mSiO2-SO3H composites). The composites have a large specific surface (173 m2·g-1), appropriate pore sizes (typically 3.7 nm), and sulfo-functionalized pore walls. Magnetic separation can be accomplished within 10 s. The unique properties of the composites allow both MET and Glim to be selectively and quickly extracted from plasma sample. Following magnetic separation and elution by 50% aqueous acetonitrile with 4% ammonium solution, the two drugs were quantified by LC-MS/MS analysis. The assay has high selectivity, good linearity (2.5-4000 ngâ¢mL-1 for MET and 0.02-1600 ngâ¢mL-1 for Glim), a low detection limit (as low as 60 pgâ¢mL-1 for MET and 1 pgâ¢mL-1 for Glim), excellent recovery (above 92.2%), and good precision (RSDs <12%). The method was successfully applied in a pharmacokinetic study in beagle dogs. Graphical abstract Schematic representation of the synthesis of sulfo-functionalized magnetic graphene/mesoporous silica composites, giving a material of type magG@mSiO2-SO3H. Results showed its great potential as a feasible and alternative adsorbent for the selective extraction of MET and Glim from complicated biological samples.
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Liposarcoma is the most common soft tissue malignancy. We investigated the relationship between the expression of fibroblast growth factor -21 protein and recurrence in the liposarcoma tissues from 40 patients. The patients were divided into two groups (low/no- and high-expressing) for further survival analysis according to fibroblast growth factor -21 expression in their tumor tissue. Immunohistochemical staining showed that fibroblast growth factor -21 protein was located in the cytoplasm. The fibroblast growth factor -21 protein was significantly less expressed in liposarcoma than in normal tissue (pâ¯<â¯0.05). Fibroblast growth factor -21 protein expression was related to gender, but not age, cell differentiation or tumor size. The patients in the low/no fibroblast growth factor 21 expression group were more likely to relapse and die in a shorter period of time. The patients in the high-expression group had a better prognosis and less recurrence. fibroblast growth factor -21 has the potential to act both as a biomarker for liposarcoma prognosis as well as a marker for the response to therapy.
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Fatores de Crescimento de Fibroblastos/análise , Lipossarcoma/diagnóstico , Biomarcadores Tumorais/análise , Feminino , Humanos , Imuno-Histoquímica , Lipossarcoma/patologia , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/diagnóstico , Recidiva Local de Neoplasia/patologia , Prognóstico , Análise de SobrevidaRESUMO
Under the conditions of the polarized light, The reflective surface of the object is affected by many factors, refractive index, surface roughness, and so the angle of incidence. For the rough surface in the different wavelengths of light exhibit different reflection characteristics of polarization, a spectral polarimetric BRDF based on Kirchhof theory is proposee. The spectral model of complex refraction index is combined with refraction index and extinction coefficient spectral model which were got by using the known complex refraction index at different value. Then get the spectral model of surface roughness derived from the classical surface roughness measuring method combined with the Fresnel reflection function. Take the spectral model of refraction index and roughness into the BRDF model, then the spectral polarimetirc BRDF model is proposed. Compare the simulation results of the refractive index varies with wavelength, roughness is constant, the refraction index and roughness both vary with wavelength and origin model with other papers, it shows that, the spectral polarimetric BRDF model can show the polarization characteristics of the surface accurately, and can provide a reliable basis for the application of polarization remote sensing, and other aspects of the classification of substances.
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Fusarium oxysporum f. sp. cubense (FOC) is the causal agent of banana Fusarium wilt and has become one of the most destructive pathogens threatening the banana production worldwide. However, few genes related to morphogenesis and pathogenicity of this fungal pathogen have been functionally characterized. In this study, we identified and characterized the disrupted gene in a T-DNA insertional mutant (L953) of FOC with significantly reduced virulence on banana plants. The gene disrupted by T-DNA insertion in L953 harbors an open reading frame, which encodes a protein with homology to α-1,6-mannosyltransferase (OCH1) in fungi. The deletion mutants (ΔFoOCH1) of the OCH1 orthologue (FoOCH1) in FOC were impaired in fungal growth, exhibited brighter staining with fluorescein isothiocyanate (FITC)-Concanavalin A, had less cell wall proteins and secreted more proteins into liquid media than the wild type. Furthermore, the mutation or deletion of FoOCH1 led to loss of ability to penetrate cellophane membrane and decline in hyphal attachment and colonization as well as virulence to the banana host. The mutant phenotypes were fully restored by complementation with the wild type FoOCH1 gene. Our data provide a first evidence for the critical role of FoOCH1 in maintenance of cell wall integrity and virulence of F. oxysporum f. sp. cubense.
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Fusarium/metabolismo , Manosiltransferases/metabolismo , Parede Celular/metabolismo , Celofane/química , DNA Bacteriano/genética , Fusarium/genética , Fusarium/patogenicidade , Hifas/genética , Hifas/metabolismo , Manosiltransferases/genética , Musa/microbiologia , Mutação , Filogenia , Raízes de Plantas/microbiologia , Esporos Fúngicos/genética , Esporos Fúngicos/metabolismo , VirulênciaRESUMO
In this case report, we present the unique and intriguing case of a 57-year-old man who experienced exertional palpitations and shortness of breath for 5 years. He was diagnosed with idiopathic heart failure three years ago, leading to diuretic treatment. Physical examination revealed notable left lower extremity swelling, severe varicose veins, and cardiac murmurs. Echocardiography showed significant cardiac enlargement and severe functional mitral and tricuspid valve regurgitation. Computed tomography (CT) imaging uncovered a 10 mm left common iliac arteriovenous fistula, causing abnormal early filling of the inferior vena cava (IVC) and marked IVC dilation. Open surgical repair of the arteriovenous fistula resulted in symptom relief and improved cardiac function. This case underscores the importance of considering unusual causes in heart failure patients and highlights the value of early diagnosis and intervention in complex cardiac-vascular interactions.
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Fístula Arteriovenosa , Derivação Arteriovenosa Cirúrgica , Insuficiência Cardíaca , Insuficiência da Valva Tricúspide , Humanos , Masculino , Pessoa de Meia-Idade , Fístula Arteriovenosa/diagnóstico por imagem , Fístula Arteriovenosa/etiologia , Ecocardiografia , Insuficiência Cardíaca/cirurgia , Insuficiência Cardíaca/complicações , Insuficiência da Valva Tricúspide/cirurgia , Veia Cava Inferior/diagnóstico por imagem , Veia Cava Inferior/cirurgiaRESUMO
Ginger, a well-known spice plant, has been used widely in medicinal preparations for pain relief. However, little is known about its analgesic components and the underlying mechanism. Here, we ascertained, the efficacy of ginger ingredient 8-Shogaol (8S), on inflammatory pain and tolerance induced by morphine, and probed the role of TRPV1 in its analgesic action using genetic and electrophysiology approaches. Results showed that 8S effectively reduced nociceptive behaviors of mice elicited by chemical stimuli, noxious heat as well as inflammation, and antagonized morphine analgesic tolerance independent on opioid receptor function. Genetic deletion of TRPV1 significantly abolished 8S' analgesia action. Further calcium imaging and patch-clamp recording showed that 8S could specifically activate TRPV1 in TRPV1-expressing HEK293T cells and dorsal root ganglion (DRG) neurons. The increase of [Ca2+]i in DRG was primarily mediated through TRPV1. Mutational and computation studies revealed the key binding sites for the interactions between 8S and TRPV1 included Leu515, Leu670, Ile573, Phe587, Tyr511, and Phe591. Further studies showed that TRPV1 activation evoked by 8S resulted in channel desensitization both in vitro and in vivo, as may be attributed to TRPV1 degradation or TRPV1 withdrawal from the cell surface. Collectively, this work provides the first evidence for the attractive analgesia of 8S in inflammatory pain and morphine analgesic tolerance mediated by targeting pain-sensing TRPV1 channel. 8S from dietary ginger has potential as a candidate drug for the treatment of inflammatory pain.
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Catecóis , Gânglios Espinais , Canais de Cátion TRPV , Zingiber officinale , Canais de Cátion TRPV/metabolismo , Zingiber officinale/química , Animais , Humanos , Células HEK293 , Gânglios Espinais/efeitos dos fármacos , Gânglios Espinais/metabolismo , Catecóis/farmacologia , Camundongos , Masculino , Camundongos Endogâmicos C57BL , Inflamação/tratamento farmacológico , Analgésicos/farmacologia , Morfina/farmacologia , Cálcio/metabolismoRESUMO
The current study aimed to determine the effects of different levels of fermented Juncao grass (FG) on growth parameters, blood constituents, immunity, and antioxidative properties of broilers. A total of 240 (21-d-old) broiler chicks were randomly distributed to four dietary treatments of sixty birds, with six replicate pens and ten birds in each. Fermented grass was added to the basal diet at four levels with 0, 5, 10, and 15% FG. The results revealed that broilers fed 5% FG had significantly higher (P < 0.05) final body weight (FBW), average daily gain (WG) and average daily feed intake (ADFI). The best conversion ratio (FCR) was recorded for broilers supplemented with 5% FG compared to the group supplemented with 15% FG (P < 0.05). Increasing FG % decreased (P < 0.05) anti-inflammatory cytokines IL-4, IL-6, and IL-10. However, FG increased (P < 0.05) proinflammatory cytokines IFN-γ, IL-1, IL-2, IL-12, and TNF-α (P < 0.05). Moreover, IgA, IgG, and IgM levels increased (P < 0.05) with increasing FG %. In addition, increasing FG % in broiler rations significantly increased (P < 0.05) serum antioxidant levels of T-AOC, GSH-PX, SOD, CAT, NO and GSH, but decreased (P < 0.05) MDA levels compared to the control group. Conclusively, fermented Juncao grass would be considered a novel herbal feed additive for improving broiler performance, immunity, antioxidant, and health status. Nevertheless, further research at the molecular level is needed to quantify the effects of these herbal components on cellular and humoral immune functions in broiler chickens.
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Antioxidantes , Galinhas , Animais , Galinhas/fisiologia , Suplementos Nutricionais , Dieta/veterinária , Citocinas , Imunidade , Ração Animal/análiseRESUMO
Morphological changes that occur during kidney injury involve actin skeleton remodeling. Here we tested whether heat-shock protein 27 (HSP27), a small stress response protein involved in cytoskeletal remodeling, protects the kidney from tubulointerstitial fibrosis in obstructive nephropathy. Tubular cell HSP27 immunostaining was significantly increased in human kidneys with ureteropelvic junction obstruction, supporting the clinical relevance of our studies. To develop an animal model for mechanistic studies, we generated transgenic mice that specifically overexpress human HSP27 in renal tubules, under the kidney androgen-regulated protein promoter, and determined the effects of HSP27 overexpression on epithelial-to-mesenchymal transition and tubulointerstitial fibrosis following unilateral ureteral obstruction. This was associated with decreased fibrogenesis as evidenced by significant declines in phosphorylated p38MAPK, collagen III, α-smooth muscle actin, 4-hydroxynonenal, and reduced trichrome staining following obstruction. Notably, E-cadherin and ß-catenin remained at the cell membrane of tubular cells in transgenic mice with an obstructed ureter. Monocyte/macrophage infiltration, however, was not significantly affected in these transgenic mice. Thus, tubular HSP27 inhibits fibrogenesis in obstructive nephropathy. Further studies are needed to determine pathways regulating the interactions between HSP27 and the E-cadherin-ß-catenin complex.
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Proteínas de Choque Térmico HSP27/metabolismo , Nefropatias/metabolismo , Nefropatias/patologia , Túbulos Renais/metabolismo , Túbulos Renais/patologia , Obstrução Ureteral/metabolismo , Obstrução Ureteral/patologia , Animais , Caderinas/metabolismo , Membrana Celular/metabolismo , Colágeno Tipo III/metabolismo , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Transição Epitelial-Mesenquimal , Feminino , Fibrose , Proteínas de Choque Térmico HSP27/genética , Humanos , Masculino , Camundongos , Camundongos Transgênicos , beta Catenina/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Fibroblast growth factor (FGF)-21 is a novel regulator of insulin-independent glucose transport in 3T3-L1 adipocytes and has glucose and triglyceride lowering effects in rodent models of diabetes. In this study, we found that FGF-21 can significantly attenuate ischemia-reperfusion (I/R) induced damage in H9c2 cells (rat heart). However, it is unclear which signal transduction pathway is involved in the cardioprotective effect of FGF-21. Thus, this study was designed to investigate the potential mechanism induced by FGF-21. The results showed that FGF-21 treatment prevented the oxidative stress and apoptosis associated with I/R damage by reducing the levels of superoxide anions, inhibiting glycogen synthase kinase (GSK) 3ß by activating Akt phosphorylation, and recovering the levels of ATP synthase pyruvate kinase isozymes M1 and protein kinase C, thereby improving energy supply. In summary, we conclude that FGF-21 protects H9c2 cells against I/R injury mainly through the Akt-GSK-3ß-caspase-3 dependent pathway, preventing oxidative stress, and recovery of the energy supply.
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Cardiotônicos , Fatores de Crescimento de Fibroblastos/farmacologia , Traumatismo por Reperfusão/tratamento farmacológico , Apoptose/efeitos dos fármacos , Western Blotting , Caspase 3/fisiologia , Contagem de Células , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Análise por Conglomerados , Eletroforese em Gel de Poliacrilamida , Etídio , Quinase 3 da Glicogênio Sintase/fisiologia , Humanos , Miócitos Cardíacos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Proteômica , Proteínas Proto-Oncogênicas c-akt/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/efeitos dos fármacos , Sincalida/metabolismo , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
BACKGROUND: To assess the outcomes of cataract surgery performed by novice surgeons during training in a rural programme. DESIGN : Retrospective study. PARTICIPANTS: Three hundred thirty-four patients operated by two trainees under supervision at rural Chinese county hospitals. METHODS: Two trainees performed surgeries under supervision. Visual acuity, refraction and examinations were carried out 3 months postoperatively. MAIN OUTCOME MEASURES: Postoperative uncorrected visual acuity, pinhole visual acuity, causes of visual impairment (postoperative uncorrected visual acuity < 6/18) RESULTS: Among 518 operated patients, 426 (82.2%) could be contacted and 334 (64.4% of operated patients) completed the examinations. The mean age was 74.1 ± 8.8 years and 62.9% were women. Postoperative uncorrected visual acuity was available in 372 eyes. Among them, uncorrected visual acuity was ≥6/18 in 278 eyes (74.7%) and <6/60 in 60 eyes (16.1%), and 323 eyes (86.8%) had pinhole visual acuity ≥ 6/18 and 38 eyes (10.2%) had pinhole visual acuity < 6/60. Main causes of visual impairment were uncorrected refractive error (63.9%) and comorbid eye disease (24.5%). Comorbid eye diseases associated with pinhole visual acuity < 6/60 (n = 23, 6.2%) included glaucoma, other optic nerve atrophy, vitreous haemorrhage and retinal detachment. CONCLUSIONS: The findings suggest that hands-on training remains safe and effective even when not implemented in centralized training centres. Further refinement of the training protocol, providing postoperative refractive services and more accurate preoperative intraocular lens calculations, can help optimize outcomes.
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Competência Clínica/normas , Educação de Pós-Graduação em Medicina , Oftalmologia/educação , Facoemulsificação/educação , Acuidade Visual/fisiologia , Idoso , Idoso de 80 Anos ou mais , China , Avaliação Educacional , Feminino , Hospitais de Condado , Hospitais Rurais , Humanos , Complicações Intraoperatórias , Masculino , Pessoa de Meia-Idade , Satisfação do Paciente , Complicações Pós-Operatórias , Refração Ocular/fisiologia , Estudos Retrospectivos , População Rural , Resultado do TratamentoRESUMO
In order to make diffraction energy of concave gratings more concentrated in the desired order, the present paper puts forward that the concave blazed grating with variable groove angles could be fabricated on the concave substrates by mechanical ruling method, and the theoretical method of simultaneously calculating the diffraction efficiency in the main section and non-main section is deduced by using Fresnel-Kirchhoff's diffraction formula, which makes up the shortage of the diffraction efficiency calculated only in the main section. Finally, the diffraction efficiency curves varied with wavelength is simulated by Matlab software, and the variation laws of the diffraction efficiency are compared for different production methods and application parameters, which provides a valuable reference for the design and production of the concave gratings.
RESUMO
Metabolic disorder is a major characteristic of cancer cells, and controlling genes involved in metabolic shifts can be an effective strategy for cancer treatment. Andrographolide (AG), a diterpenoid lactone, is widely recognized as a natural anticancer drug due to its ability to inhibit cancer growth. The present study aimed to investigate the mechanism underlying the mitochondrialmediated anticancer effect of AG by inhibiting pyruvate dehydrogenase kinase 1 (PDK1) expression in lung cancer cells. Cells were treated with AG and PDK1 mRNA and protein expression was determined using reverse transcriptionquantitative PCR and western blotting, respectively. As a result, AG significantly inhibited the viability of human lung cancer cells and suppressed aerobic glycolysis by decreasing lactate generation. AG further decreased the PDK1 protein and mRNA levels in a dosedependent manner. AGinduced cell death was assessed by flow cytometry and fluorescence microscopy. AG induced apoptotic cell death that was associated with the cleavage of poly (ADP ribose) polymerase, activation of caspase3, and mitochondrial damage, which was associated with an increase in reactive oxygen species and loss of mitochondrial membrane potential. AGinduced cell death was partially suppressed via PDK1 overexpression in lung cancer cells. Therefore, the anticancer effects of AG on human lung cancer cells may negatively regulate the expression of PDK1.
Assuntos
Diterpenos , Neoplasias Pulmonares , Humanos , Piruvato Desidrogenase Quinase de Transferência de Acetil/metabolismo , Proteínas Serina-Treonina Quinases/genética , Apoptose , Diterpenos/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Glicólise , Linhagem Celular Tumoral , Proliferação de CélulasRESUMO
Depression is a common mental disorder with an increasing incidence. Several studies have demonstrated that cortical DNA hypomethylation is associated with depression-like behaviors. This study aims to investigate whether maternal vitamin D deficiency (VDD) induces depression-like behaviors and to explore the effects of folic acid supplement on VDD-induced cortical DNA hypomethylation in adult offspring. Female mice were fed with a VDD diet, beginning at 5 weeks of age and throughout pregnancy. Depression-like behaviors were evaluated, and cortical 5-methylcytosine (5mC) content was detected in adult offspring. Results showed that depression-like behaviors were observed in adult offspring of the VDD group. Cortical Ache and Oxtr mRNAs were upregulated in female offspring of the VDD group. Cortical Cpt1a and Htr1b mRNAs were increased in male offspring of the VDD group. Moreover, cortical 5mC content was reduced in offspring of VDD-fed dams. The additional experiment showed that serum folate and cortical S-adenosylmethionine (SAM) contents were decreased in the offspring of the VDD group. Folic acid supplement attenuated VDD-induced SAM depletion and reversed cortical DNA methylation. Moreover, folic acid supplement attenuated VDD-induced upregulation of depression-related genes. In addition, folic acid supplement alleviated maternal VDD-induced depression-like behaviors in adult offspring. These results suggest that maternal VDD induces depression-like behavior in adult offspring by reducing cortical DNA methylation. The gestational folic acid supplement prevents VDD-induced depression-like behavior by reversing cortical DNA hypomethylation in adult offspring.
Assuntos
Ácido Fólico , Deficiência de Vitamina D , Gravidez , Animais , Masculino , Feminino , Camundongos , Ácido Fólico/farmacologia , Metilação de DNA , Depressão/etiologia , Depressão/prevenção & controle , DNARESUMO
We report here a sensitive and environmentally benign technique for the visualization of DNA by using eosin Y, which appeared as a transparent and colorless band under the opaque gel matrix background. As low as 0.2-0.6 ng of DNA can be visualized within 1 h, similar to that of the most commonly used ethidium bromide stain. Furthermore, the mechanism of the newly developed method was also described. According to the results, it can be concluded that the complex between DNA and eosin Y is mainly formed by the function of groove binding, but not intercalate itself into the double helix structure of DNA.