RESUMO
Low-tannin sorghum is an excellent energy source in pig diets. However, sorghum contains several anti-nutritional factors that may have negative effects on nutrient digestibility. The impacts of proteases on growth performance, nutrient digestibility, blood parameters, and gut microbiota of growing pigs fed sorghum-based diets were studied in this study. Ninety-six pigs (20.66 ± 0.65 kg BW) were allocated into three groups (eight pens/group, four pigs/pen): (1) CON (control diet, sorghum-based diet included 66.98% sorghum), (2) PRO1 (CON + 200 mg/kg proteases), (3) PRO2 (CON + 400 mg/kg proteases) for 28 d. No differences were observed in growth performance and apparent total tract digestibility (ATTD) of nutrients between CON and PRO1 groups. Pigs fed PRO2 diet had increased (P < 0.05) BW on d 21 and 28, and increased (P < 0.05) average daily gain during d 14-21 and the overall period compared with pigs fed CON diet. In addition, pigs fed PRO2 diet had improved (P < 0.05) ATTD of gross energy, CP, and DM compared with pigs fed CON and PRO1 diets. Pigs fed PRO2 diet had lower (P < 0.05) plasma globulin (GLB) level and higher (P < 0.05) plasma glucose, albumin (ALB) and immunoglobulin G levels, and ALB/GLB ratio than pigs fed CON and PRO1 diets. Furthermore, pigs fed PRO2 diet had decreased (P < 0.05) the relative abundance of Acidobacteriota at the phylum level and increased (P < 0.05) the relative abundance of Prevotella_9 at the genus level. The linear discriminant analysis effect size analysis also showed that pigs fed PRO2 diet had significantly enriched short-chain fatty acid-producing bacteria, such as Subdoligranulum and Parabacteroides. In conclusion, protease supplementation at 400 mg/kg improved the growth performance of growing pigs fed sorghum-based diets, which may be attributed to the improvement of nutrient digestibility, host metabolism, immune status and associated with the altered gut microbiota profiles.
Assuntos
Microbioma Gastrointestinal , Sorghum , Animais , Suínos , Peptídeo Hidrolases , Digestão , Ração Animal/análise , Dieta/veterinária , Nutrientes , Suplementos Nutricionais/análise , Fenômenos Fisiológicos da Nutrição AnimalRESUMO
OBJECTIVE: The aim of this study was to investigate the modular characteristics and mechanism of action of Chinese herbs for vascular calcification (VC) treatment. MATERIALS AND METHODS: Network pharmacology coupled with literature data mining was utilized to assess the Chinese herbal clinical performance as well as its similarity, characteristics, ingredient, target, and Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and network construction. RESULTS: The top 15 medications from the literature, according to the usage, and 190 active chemicals, 183 common targets between medication and VC-related targets were weeded out. Analysis of the relationships between the active ingredients, pharmacological targets, and signaling pathways helped to clearly define the therapeutic effect of Traditional Chinese Medicine (TCM). Importantly, we discovered seven most hub proteins (AKT1, CTNNB1, TNF, EGFR, TP53, JUN and IL-6) and two of the herbs' most fundamental ingredients (Formononetin and Luteolin) in TCM-mediated VC suppression. Mechanistically, the metabolic pathways [AGE-RAGE pathway, interleukin-17 (IL-17) pathway, and p53 pathway] as well as smooth muscle adaptation (functional remodeling) and oxidoreductase activity (redox homeostasis modulating) are also crucially implicated. CONCLUSIONS: Our work, accomplished by network pharmacology and data mining, increases our understanding of TCM in VC therapy and may offer insightful information for future drug discovery investigations.
Assuntos
Medicamentos de Ervas Chinesas , Calcificação Vascular , Humanos , Medicina Tradicional Chinesa , Farmacologia em Rede , Calcificação Fisiológica , Mineração de Dados , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Simulação de Acoplamento MolecularRESUMO
To seek viable alternatives to antibiotics, we determined the combinatorial effects of Lactobacillus and a quorum quenching enzyme (QQE) on broiler growth performance, antioxidant capacity, immune responses, and cecal microbial populations. In total, 360 one-day-old male broilers (Ross 308) were randomly allotted to 3 dietary treatments, with 12 replicate pens/treatment and 10 birds/replicate pen. Dietary treatments lasted 42 d and comprised: corn-soybean meal basal diet (control group, CON); control plus antibiotic growth promoter supplement group (AGP); and control plus Lactobacillus and QQE supplement group (LQ). Dietary LQ supplementation significantly increased final body weight (BW) and average daily gain (ADG) when compared with CON and AGP groups between 22 and 42 d and 1 to 42 d (P < 0.05). No significant differences were observed for serum superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) levels between treatments (P > 0.05). A higher concentration of total antioxidant capacity (T-AOC) was observed on d 42 in the LQ group (P = 0.06). Feeding LQ significantly increased serum immunoglobulins (IgA and IgG) levels when compared with other treatments (P < 0.05). A statistical trend was also observed for increased cecal butyrate levels (P = 0.06) in the LQ group. Bacterial α-diversity was unaffected by dietary treatments (P > 0.05). However, from principal component analysis (PCoA), the microbial community structure was different between the LQ and AGP groups. Diet supplemented with LQ significantly (P < 0.05) decreased the relative abundance of Synergistota and Proteobacteria and significantly (P < 0.05) increased the proportion of Ruminococcaceae and Faecalibacterium. Thus, supplemental LQ improved growth performance, immune status, and modulated intestinal microbial communities in broilers. We provide a new perceptive on antibiotic substitutes in the poultry industry.
Assuntos
Antioxidantes , Microbioma Gastrointestinal , Lactobacillus , Percepção de Quorum , Ração Animal/análise , Animais , Antibacterianos/farmacologia , Antioxidantes/farmacologia , Galinhas/fisiologia , Dieta/veterinária , Suplementos Nutricionais/análise , Imunidade , MasculinoRESUMO
Limited therapeutic options are available for treating deep caries. Those materials with potential of a dual effect to remineralize hard tissue and regenerate defective dentin tissues could be used as a new strategy for deep caries treatment. However, the application of the single component remains a challenge mainly because they lack calcium and phosphorus, are easily degraded, and are difficult to retain in the intricate body fluid environment. Considering the abundant source of calcium and phosphorus as well as the delivery performance of mesoporous bioactive glass (MBG), an amelogenin-derived peptide (QP5), which has a significant role in hard tissue remineralization, was loaded to fabricate a novel composite. After the synthesis of highly ordered MBG using a sol-gel method, the QP5 peptide was loaded increasingly by its extensive porous structure and enhanced electrostatic absorption. When used in an acidic environment, the MBG/QP5 composite presented pH-responsiveness, releasing therapeutic ions and functional peptides in a sequential cascade, and eventually adjusted the pH to a neutral state. The composite was internalized by dental pulp cells through a clathrin-mediated pathway and influenced by cell membrane lipid raft regulation. It could be also transported through the macro-pinocytotic pathway. Compared to the single treatment of peptide QP5 in 48 h, the composite facilitated a higher level of retention of the intracellular peptides. The composite further promoted migration and odontogenesis of dental pulp cells, including the improved activity of alkaline phosphatase, increased formation of mineralized nodules, and upregulated expression of mineralization-related genes compared to using MBG or QP5 alone. The composite further induced the dentin-like layer in a rat pulp capping model. The results suggested that this intelligent material with pH-responsiveness provides a promising alternative treatment method for biomimetic restoration of deep caries.
Assuntos
Materiais Biocompatíveis , Alicerces Teciduais , Animais , Materiais Biocompatíveis/farmacologia , Cálcio , Endocitose , Vidro/química , Odontogênese , Peptídeos , Fósforo , Porosidade , Ratos , Alicerces Teciduais/químicaRESUMO
The complexity of burn metabolism and limitations of its cognition are the core factors leading to dissatisfactory efficacy of nutritional therapy in severe burn patients. Precise understanding of burn metabolic rules is a prerequisite for improving the pertinence and effectiveness of nutritional therapy. The previous division of burn metabolic stages based on energy consumption did not pay enough attention to substance metabolism, so it is difficult to fully comprehend the overall change pattern and stage characteristics of burn metabolism. Through the effective integration of the metabonomics, physiomics, and clinical data, combining the pathophysiological characteristics of burns at different clinical phases, this paper suggested that the metabolism of severe burns should be divided into four stages and proposed the corresponding nutritional strategies and measures according to different metabolic characteristics, in order to provide a basis for further enhancing the nutritional efficacy in burn patients.
Assuntos
Queimaduras , Queimaduras/terapia , Humanos , Metabolômica , Apoio NutricionalRESUMO
Objective: To evaluate the clinical efficacy of dietary supplement Licofor in the treatment of dry eye associated with meibomian gland dysfunction (MGD). Methods: This was a prospective, randomized controlled clinical trial. Sixty patients [25 males, 35 females, aged (42±13) years] who had dry eye associated with MGD were recruited in Xiangya Hospital of Central South University from December 2018 to October 2019. The patients were equally divided into two groups: 30 cases (60 eyes) in the experimental group and 30 cases (60 eyes) in the control group. All subjects were treated with eye hot compress, artificial tears and antibiotic ointment. After that, the experimental group and control group were received dietary supplementary Licofor or placebo daily for 12 weeks. The symptoms and signs of dry eye, morphology and function of meibomian gland, and inflammatory response were assessed at the beginning, 4th, 8th and 12th week of treatment. Results: After 12 weeks of treatment, statistically significant improvements in ocular surface disease index (OSDI) scores, tear break-up time (TBUT), corneal fluorescein staining (CFS), the morphology of eyelid margin, meibomian gland orifice, meibomian gland expressibility, meibum quality, and periglandular inflammatory cell density were determined in both groups (all P<0.05). In the Licofor group, the improvement of OSDI scores [16.7 (12.5, 20.8) vs 20.8 (18.8, 22.9), P<0.001], the morphology of eyelid margin, meibomian gland orifice and periglandular inflammatory cell density [443 (318, 513) vs 553 (415, 676)/mm2, P=0.002] were more significant (all P<0.05). Conclusion: The combined treatment of licofor and conventional treatment can significantly improve symptoms of dry eye, the morphology of eyelid margin, meibomian gland orifice, meibum quality, and eyelid inflammation response of dry eye associated with MGD.
Assuntos
Síndromes do Olho Seco , Doenças Palpebrais , Disfunção da Glândula Tarsal , Suplementos Nutricionais , Síndromes do Olho Seco/tratamento farmacológico , Doenças Palpebrais/tratamento farmacológico , Feminino , Humanos , Masculino , Glândulas Tarsais , Estudos Prospectivos , Lágrimas , Resultado do TratamentoRESUMO
This study was to evaluate the effects of supplementation of AA form (crystalline vs. protein bound) in low-protein diets on growth, metabolic, and immunological characteristics of pigs. A total of 80 barrows (PIC 327 × 1050; 15.57 ± 0.13 kg BW and 48 ± 2 d of age), housed in 4 pigs per pen with 5 pens per treatment, were assigned to 4 dietary treatments of 17, 15, and 13% CP and 13% CP plus casein for 28 d. The crystalline AA were supplemented to meet the requirement of indispensable AA in pigs. Results showed that pigs fed the 13% CP diet or the 13% CP plus casein diet had lower ( < 0.01) ADG and ADFI and a greater ( < 0.01) feed:gain ratio than pigs fed the 17% CP or 15% CP diets over the 4-wk study period. Compared with other diets, pigs fed the 13% CP diet had decreased concentrations of plasma urea nitrogen, albumin ( < 0.01), and mRNA expressions of Toll-like receptor 4 (), nuclear factor kappa B (; < 0.05), and Toll-interacting protein (; < 0.01) in the ileum and also increased activity of plasma glutamate-pyruvate transaminase ( < 0.05) and concentrations of IL-1ß ( < 0.05) and tumor necrosis factor-α ( < 0.01); however, these characteristics were partly normalized by feeding the 13% CP plus casein diet. Furthermore, the plasma concentration of insulin-like growth factor 1 (IGF-1; < 0.01) and mRNA expressions of protein kinase B (), mammalian target of rapamycin (), and ribosomal protein S6 kinase () in longissimus muscle were increased ( < 0.05) in pigs fed the 13% CP plus casein diet relative to pigs fed the 17% CP or 15% CP diets. In summary, reducing dietary CP level from 17% to 15% had no effect on growth, metabolic, and immunological characteristics of 15- to 35-kg pigs. A further reduction of dietary CP level up to 13% would lead to poor growth performance, but metabolic and immunological characteristics were partly normalized using protein-bound AA to replace synthesized AA in the 13% CP diet.
Assuntos
Aminoácidos/administração & dosagem , Dieta com Restrição de Proteínas/veterinária , Suplementos Nutricionais , Suínos/fisiologia , Ração Animal/análise , Animais , Nitrogênio da Ureia Sanguínea , Proteínas Alimentares/administração & dosagem , Íleo/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Masculino , NF-kappa B/genética , Albumina Sérica , Suínos/crescimento & desenvolvimento , Suínos/imunologia , Receptor 4 Toll-Like/genética , Fator de Necrose Tumoral alfa/genéticaRESUMO
BACKGROUND: Treatment failure of prostate cancer (PCa) is often due to bone metastasis. Celastrol, an active constituent of Tripterygium wilfordii roots, has shown anti-tumor effects in previous studies in accordance with its indication in traditional Chinese medicine. METHODS: Using a PC-3 cell model, in vitro assays were performed to evaluate the effects of celastrol on proliferation, migration (wound healing assay), tissues invasion (Transwell-Matrigel penetration assay) and vascular endothelial growth factor (VEGF) secretion (enzyme-linked immunosorbent assay). An intra-tibia injection mouse model was used to assess the effect of celastrol on PCa bone metastasis in vivo. RESULTS: Pretreatment with celastrol significantly reduced proliferation of PC-3 cells in a dose-dependent manner and cell migration was much slower than in controls. Significantly fewer cells penetrated the gel-membrane after celastrol administration and their skeletal invasive ability was significantly reduced in a dose-dependent manner. Correspondingly, a significant, dose-dependent decrease in VEGF secretion was observed. In the in vivo mouse model, pretreatment with celastrol (8 µmol l-1) inhibited the tumorigenicity of PC-3 cells so that almost no bone invasion occurred as compared with control injections. Histological examinations using hematoxylin and eosin staining showed that tibiae injected with celastrol pretreated PC-3 cells retained their natural bone structure. CONCLUSIONS: Celastrol may have preventive potential against PCa bone metastasis.
Assuntos
Neoplasias Ósseas/tratamento farmacológico , Medicina Tradicional Chinesa , Neoplasias da Próstata/tratamento farmacológico , Triterpenos/administração & dosagem , Neoplasias Ósseas/genética , Neoplasias Ósseas/patologia , Neoplasias Ósseas/secundário , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Invasividade Neoplásica/patologia , Metástase Neoplásica , Triterpenos Pentacíclicos , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Triterpenos/química , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
The objective of this study was to determine if a moderate or high reduction of dietary CP, supplemented with indispensable amino acids (IAA), would affect growth, intestinal morphology and immunological parameters of pigs. A total of 40 barrows (initial BW=13.50±0.50 kg, 45±2 day of age) were used in a completely randomized block design, and allocated to four dietary treatments containing CP levels at 20.00%, 17.16%, 15.30% and 13.90%, respectively. Industrial AA were added to meet the IAA requirements of pigs. After 4-week feeding, blood and tissue samples were obtained from pigs. The results showed that reducing dietary CP level decreased average daily gain, plasma urea nitrogen concentration and relative organ weights of liver and pancreas (P<0.01), and increased feed conversion ratio (P<0.01). Pigs fed the 13.90% CP diet had significantly lower growth performance than that of pigs fed higher CP at 20.00%, 17.16% or 15.30%. Moreover, reducing dietary CP level decreased villous height in duodenum (P<0.01) and crypt depth in duodenum, jejunum and ileum (P<0.01). The reduction in the dietary CP level increased plasma concentrations of methionine, alanine (P<0.01) and lysine (P<0.05), and decreased arginine (P<0.05). Intriguingly, reducing dietary CP level from 20.00% to 13.90% resulted in a significant decrease in plasma concentration of IgG (P<0.05), percentage of CD3+T cells of the peripheral blood (P<0.01), also down-regulated the mRNA abundance of innate immunity-related genes on toll-like receptor 4, myeloid differentiation factor 88 (P<0.01) and nuclear factor kappa B (P<0.05) in the ileum. These results indicate that reducing dietary CP level from 20.00% to 15.30%, supplemented with IAA, had no significant effect on growth performance and had a limited effect on immunological parameters. However, a further reduction of dietary CP level up to 13.90% would lead to poor growth performance and organ development, associated with the modifications of intestinal morphology and immune function.
Assuntos
Aminoácidos/farmacologia , Dieta com Restrição de Proteínas/veterinária , Proteínas Alimentares/administração & dosagem , Suplementos Nutricionais , Intestinos/anatomia & histologia , Suínos/crescimento & desenvolvimento , Suínos/imunologia , Aminoácidos/administração & dosagem , Aminoácidos/sangue , Aminoácidos/metabolismo , Ração Animal/análise , Animais , Nitrogênio da Ureia Sanguínea , Peso Corporal , Proteínas Alimentares/farmacologia , Duodeno/anatomia & histologia , Íleo/anatomia & histologia , Imunoglobulina G/sangue , Jejuno/anatomia & histologia , Tamanho do Órgão , Suínos/anatomia & histologia , Suínos/sangue , Linfócitos T/citologia , Receptor 4 Toll-Like/genética , Aumento de Peso/efeitos dos fármacosRESUMO
BACKGROUND AND PURPOSE: Insulin secretion from isolated pancreatic islets is a pivotal assay in developing novel insulin secretagogues, given its good correlation with in vivo efficacy. Because the supply of human islets is limited, this assay is typically run with rodent islets, which do not address species differences and are low-throughput, because of the size matching or volume normalization required. Here we have evaluated the suitability of human re-aggregated islets for this assay. EXPERIMENTAL APPROACH: We generated re-aggregated human islets of a consistent size, using micromolds and compared their responses with those of native human and rat islets, to known secretagogues and inhibitors of insulin release. KEY RESULTS: Insulin secretion from rat islets, human islets and human re-aggregated cell clusters was concentration-dependently increased by glucose. The calcium channel agonist, Bay K 8644, stimulated insulin secretion in native rat islets and human re-aggregated islets, but not native human islets. Glibenclamide and tolbutamide were more effective and potent in re-aggregated human clusters compared with the other two preparations. Rat islets outperformed both human preparations of islets in response to caffeine, carbachol and glucagon-like peptide-1. Re-aggregated human islet clusters were more sensitive to somatostatin, diazoxide and sodium azide, but rodent islets were more sensitive to nifedipine. CONCLUSIONS AND IMPLICATIONS: Human re-aggregated clusters of islet cells, of a constant size were more responsive to all compounds tested than native human islets. Importantly, the assay variability was less in the re-aggregated cluster preparations, which suggests that such re-aggregated cells could be useful for drug development.
Assuntos
Comunicação Celular , Avaliação Pré-Clínica de Medicamentos/métodos , Hipoglicemiantes/farmacologia , Insulina/metabolismo , Ilhotas Pancreáticas/efeitos dos fármacos , Adulto , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Glucose/metabolismo , Humanos , Secreção de Insulina , Ilhotas Pancreáticas/metabolismo , Masculino , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Especificidade da EspécieRESUMO
We previous identified the antifibrotic active ingredients from Carapax Trionycis as two peptides. Here, we synthesized these two peptides (peptide 1 and peptide 2) by a solid phase method and examined their effects on proliferation and activation of cultured hepatic stellate cells (HSC) which are the main ECM (extracellular matrix)-producing cells in fibrosis progression. We demonstrated that peptide 1 and peptide 2 significantly reduced HSC proliferation and activation in a dose dependent manner. Further, peptide 1 and peptide 2 could interfere with TGF-signaling by down-regulating Smad 3 phosphorylation. Thus, these synthetic peptides of Carapax Trionycis could inhibit proliferation and activation of HSC and might be used as a candidate for treatment of liver fibrosis.
Assuntos
Exoesqueleto , Células Estreladas do Fígado/efeitos dos fármacos , Medicina Tradicional Chinesa , Proteínas de Répteis/farmacologia , Fator de Crescimento Transformador beta1/antagonistas & inibidores , Tartarugas , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Matriz Extracelular/metabolismo , HumanosRESUMO
"Total petroleum hydrocarbons" (TPHs) or "petroleum hydrocarbons" (PHCs) are one of the most widespread soil pollutants in Canada, North America, and worldwide. Clean-up of PHC-contaminated soils and sediments costs the Canadian economy hundreds of million of dollars annually. Much of this activity is driven by the need to meet regulated levels of PHC in soil. These PHC values are legally required to be assessed using standard methods. The method most commonly used in Canada, specified by the Canadian Council of Ministers of the Environment (CCME), measures the total hydrocarbon concentrations in a soil by carbon range (Fraction 1: C(6)-C(10); Fraction 2: C(10)-C(16), Fraction 3: C(16)-C(34): and Fraction 4: C(34)+). Using the CCME method, all of the materials extractible by a mixture of 1:1 hexane:acetone are considered to be petroleum hydrocarbon contaminants. Many hydrocarbon compounds and other extractible materials in soil, however, may originate from non-petroleum sources. Biogenic organic compounds (BOCs) is a general term used to describe a mixture of organic compounds, including alkanes, sterols and sterones, fatty acids and fatty alcohols, and waxes and wax esters, biosynthesized by living organisms. BOCs are also produced during the early stages of diagenesis in recent aquatic sediments. BOC sources could include vascular plants, algae, bacteria and animals. Plants and algae produce BOCs as protective wax coating that are released back into the sediment at the end of their life cycle. BOCs are natural components of thriving plant communities. Many solvent-extraction methods for assessing soil hydrocarbons, however, such as the CCME method, do not differentiate PHCs from BOCs. The naturally occurring organics present in soils and wet sediments can be easily misidentified and quantified as regulated PHCs during analysis using such methods. In some cases, biogenic interferences can exceed regulatory levels, resulting in remediation of petroleum impacts that are not actually present. Consequently, reliance on these methods can trigger unnecessary and costly remediation, while also wasting valuable landfill space. Therefore, it is critically important to develop new protocols to characterize and differentiate PHCs and BOCs in contaminated sediments. In this study, a new reliable gas chromatography-mass spectrometry (GC-MS) method, in combination with a derivatization technique, for characterization of various biogenic compounds (including biogenic alkanes, sterols, fatty acids and fatty alcohols) and PHCs in the same sample has been developed. A multi-criteria approach has been developed to positively identify the presence of biogenic compounds in soil and sediment samples. More than thirty sediment samples were collected from city stormwater management (SWM) ponds and wetlands across Canada. In these wet sediment samples, abundant biogenic n-alkanes, thirteen biogenic sterols, nineteen fatty carboxylic acids, and fourteen fatty alcohols in a wide carbon range have been positively identified. Both PHCs and BOCs in these samples were quantitatively determined. The quantitation data will be used for assessment of the contamination sites and toxicity risks associated with the CCME Fraction 3 hydrocarbons.
Assuntos
Cromatografia Gasosa-Espectrometria de Massas/métodos , Sedimentos Geológicos/química , Hidrocarbonetos/análise , Petróleo/análise , Poluentes do Solo/análise , Alcanos/análise , Calibragem , Ácidos Carboxílicos/análise , Ácidos Graxos/análise , Álcoois Graxos/análise , Ionização de Chama , Hidrocarbonetos/isolamento & purificação , Modelos Moleculares , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Esteróis/análiseRESUMO
Bulbus Fritillariae (BF) is the most commonly used antitussive herb in China. There are nine species of Fritillaria recorded as the drug BF in the Chinese Pharmacopoeia. Bulbus Fritillariae cirrhosae (BF cirrhosae) is a group that includes four species of BF; these four species come from wild sources with higher efficiency and lower toxicity compared to the other five species of BF. Due to reasons of carelessness and reduced costs, the other five species are often sold as BF cirrhosae. Analysis through appearance, microscopic and chemical techniques has limitations. Identifying botanical resources is a primary step in the standardization of herbal medicine. In the present article, the internal transcribed spacer 1 (ITS1) regions of the nuclear ribosomal DNA (nrDNA) of nine species and one variety of Fritillaria genus have been sequenced. A mutation site in the ITS1 region among BF cirrhosae and other species of BF has been found and can be recognized by the restriction endonuclease SmaI. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the nuclear ribosomal ITS1 region was used to differentiate BF cirrhosae from other species of BF and is a successful method in distinguishing the subgroups.
Assuntos
Antitussígenos/química , Medicamentos de Ervas Chinesas/química , Fritillaria/genética , Fitoterapia , Sequência de Bases , Primers do DNA , DNA de Plantas/análise , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de RestriçãoRESUMO
The progesterone receptor (PR) gene is expressed in cells of the anterior pituitary and hypothalamus, and PR levels are regulated by estrogen (E) in a tissue-specific fashion. To demonstrate that E induces transcription via the PR promoter, and to identify sequences within the PR promoter responsible for tissue-specific and hormonal regulation, we have utilized a defective herpes simplex virus vector for direct gene transfer into the rat pituitary and brain. We designed a viral amplicon expressing the beta-galactosidase gene under the regulation of a 2.1-kb PR promoter fragment to create a defective viral vector for gene transfer into the brain. Following injection of this vector into the pituitary and brain, its pattern of expression and ability to respond to estradiol 3-benzoate (EB) were examined. In the pituitary, lacZ activity was observed in cells of the anterior lobe (AL). However, no activity was seen in the neurointermediate lobe (NIL), demonstrating tissue specific transcriptional regulation. A approximately sixfold increase in cells demonstrating beta-galactosidase activity was observed in the AL following treatment with EB. Likewise, injection of defective viral vector into the hypothalamus followed by treatment with EB resulted in a approximately eightfold increase in cells demonstrating beta-galactosidase activity including the very cell groups responsible for EB-dependent reproductive behavior. In contrast, no vector dependent activity was observed in the caudate nucleus, a tissue with no endogenous expression of PR, despite polymerase chain reaction evidence demonstrating the presence of the vector in this tissue. These results demonstrate that the 2.1-kb PR promoter fragment contains the sequence information required for correct tissue and hormonal regulation of PR.
Assuntos
Estrogênios/metabolismo , Regulação da Expressão Gênica/genética , Hipotálamo/metabolismo , Adeno-Hipófise/metabolismo , Regiões Promotoras Genéticas/genética , Receptores de Progesterona/genética , Animais , Estrogênios/farmacologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Técnicas de Transferência de Genes , Genes Reporter/efeitos dos fármacos , Genes Reporter/genética , Vetores Genéticos/genética , Hipotálamo/efeitos dos fármacos , Adeno-Hipófise/efeitos dos fármacos , Regiões Promotoras Genéticas/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Ativação Transcricional/efeitos dos fármacos , Ativação Transcricional/genética , beta-Galactosidase/genéticaRESUMO
Progesterone receptors play a central role in neuroendocrine and behavioural regulation. To gain insight into the sex- and tissue-specific regulation of progesterone receptors, protein binding on a progesterone receptor-oestrogen response element and mRNA levels for progesterone receptor (PR)-A and PR-B were compared between female and male rats following oestradiol benzoate replacement treatment in hypothalamic and pituitary tissue. Both male and female pituitary protein extracts demonstrated an increase in nuclear protein binding activity to a progesterone receptor-oestrogen response element following oestradiol benzoate treatment. However, there was a greater difference in total binding activity seen in the female pituitary extracts compared to male pituitary protein extracts. In both cases, reflecting the binding data, oestradiol benzoate pretreatment led to an increase in pituitary PR-B messenger RNA, although this increase was significantly larger in females than in males. Oestradiol benzoate treatment also led to a significant increase in specific binding of hypothalamic nuclear proteins to the progesterone receptor oestrogen response element from both females and male hypothalamic extracts. In addition, PR-B messenger RNA was induced by oestradiol benzoate treatment in the female rat hypothalamus, under circumstances where no PR-A could be detected. The male also demonstrated an increase in PR-B messenger RNA following oestradiol benzoate treatment, with undetectable levels of PR-A, although to a lesser degree than that seen in the female. The predominance of PR-B over PR-A messenger RNA in rat hypothalamus and pituitary, and the quantitative differences between female and male rats, could both contribute to the greater responsiveness of female rats to progesterone with respect to control over luteinizing hormone release from the pituitary, and lordosis behaviour regulated by hypothalamic neurones.
Assuntos
Estradiol/farmacologia , Hipotálamo/metabolismo , Hipófise/metabolismo , RNA Mensageiro/metabolismo , Receptores de Progesterona/genética , Caracteres Sexuais , Animais , Feminino , Humanos , Masculino , Ratos , Ratos Sprague-Dawley , Útero/metabolismoRESUMO
We have reported that liver-specific deletion of IGF-I in mice (LI-IGF-I-/-) results in decreased circulating IGF-I and increased GH levels. In the present study, we determined how elimination of hepatic IGF-I modifies the hypothalamic-pituitary GH axis to enhance GH secretion. The pituitary mRNA levels of GH releasing factor (GHRF) receptor and GH secretagogue (GHS) receptor were increased in LI-IGF-I-/- mice, and in line with this, their GH response to ip injections of GHRF and GHS was increased. Expression of mRNA for pituitary somatostatin receptors, hypothalamic GHRF, somatostatin, and neuropeptide Y was not altered in LI-IGF-I-/- mice, whereas hypothalamic IGF-I expression was increased. Changes in hepatic expression of major urinary protein and the PRL receptor in male LI-IGF-I-/- mice indicated an altered GH release pattern most consistent with enhanced GH trough levels. Liver weight was enhanced in LI-IGF-I-/- mice of both genders. In conclusion, loss of liver-derived IGF-I enhances GH release by increasing expression of pituitary GHRF and GHS receptors. The enhanced GH release in turn affects several liver parameters, in line with the existence of a pituitary-liver axis.
Assuntos
Hormônio do Crescimento/metabolismo , Fator de Crescimento Insulin-Like I/fisiologia , Fígado/metabolismo , Hipófise/metabolismo , Animais , Feminino , Hormônio do Crescimento/sangue , Hormônio do Crescimento/genética , Hormônio Liberador de Hormônio do Crescimento/farmacologia , Hipotálamo/metabolismo , Fator de Crescimento Insulin-Like I/análise , Fator de Crescimento Insulin-Like I/genética , Fígado/anatomia & histologia , Masculino , Camundongos , Camundongos Knockout/genética , Neuropeptídeos/fisiologia , Tamanho do Órgão , Proteínas/genética , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/fisiologia , Receptores da Prolactina/genéticaRESUMO
A glycoconjugate with pronounced immunoactivity, designated as LbGp2, was isolated from the fruit of Lycium barbarum L. and purified to homogeneity by gel-filtration. Its carbohydrate content is up to 90.71% composed of Ara, Gal and amino acids. The molecular weight is 68.2 kDa as determined by size exclusive chromatography (SEC). The complete structure of the repeat unit of the glycan of LbGp2 was elucidated based on glycosidic linkage analysis, total acid hydrolysis, partial acid hydrolysis, 1H and 13C NMR spectroscopy. According to the experiments, the glycan possesses a backbone consisting of (1-->6)-beta-galactosyl residues, about fifty percent of which are substituted at C-3 by galactosyl or arabinosyl groups and the major nonreducing end being made of Ara (1 -->.
Assuntos
Glicoconjugados/química , Plantas Medicinais/química , Polissacarídeos/química , Solanaceae/química , Configuração de Carboidratos , Sequência de Carboidratos , China , Glicoconjugados/isolamento & purificação , Hidrólise , Dados de Sequência Molecular , Ressonância Magnética Nuclear Biomolecular , Polissacarídeos/isolamento & purificaçãoRESUMO
AIM: To isolate and purify a glycoconjugate (LbGp2) from the fruit of Lycium barbarum L. and study its immunoactivity and antioxidative activity. METHODS: By means of gel permeation chromatography, LbGp2 was purified. Based on HPLC, CE, GC, SEC and component analysis and so on, its physico-chemical properties were studied. RESULTS: Molecular weights of LbGp2 was 68.2 ku and its carbohydrate content was up to 90.7%. Component analysis showed that it composed of Ara and Gal in a molar ratio of 3:4, and 18 kinds of amino acids. The immunologic function and bioactivity of Lbp2 has been studied preliminarily. Lbp2 was shown to increase rate of phagocyticaction and phagocytic index, promote lymphocyte translation and accelerate the production of serum hemolysin. LbGp2 has distinct effect of antioxidation and the superoxide anion produced by DMSO-NaOH system was scavenged effectively. CONCLUSION: LbGp2 was shown to be a kind of homogeneous glycoconjugate with good immunoactivity and antioxidative activity.
Assuntos
Adjuvantes Imunológicos/farmacologia , Glicoconjugados/farmacologia , Lycium/química , Macrófagos Peritoneais/efeitos dos fármacos , Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/isolamento & purificação , Animais , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Feminino , Frutas/química , Glicoconjugados/química , Glicoconjugados/isolamento & purificação , Ativação Linfocitária/efeitos dos fármacos , Masculino , Camundongos , Fagocitose/efeitos dos fármacos , Plantas Medicinais/químicaRESUMO
Concentrations of selenium (Se), boron (B), and germanium (Ge) were determined in scalp hair of children with Kashin-Beck disease (KBD), in healthy children in KBD-disease endemic areas, and in healthy children in non-KBD areas. Mean Se, B, and Ge concentrations were low in children with KBD; in hair of healthy children in KBD areas, Se levels were normal but B and Ge levels were lower than in KBD-free areas. The hair levels of B and Ge were unaffected by selenium supplementation. It is suggested that B and Ge deficiency may be contributing factors in the etiology of KBD.
Assuntos
Boro/deficiência , Germânio/deficiência , Osteoartrite/etiologia , Selênio/deficiência , Adolescente , Boro/metabolismo , Criança , Suplementos Nutricionais , Feminino , Germânio/metabolismo , Cabelo/metabolismo , Mãos/diagnóstico por imagem , Humanos , Masculino , Radiografia , Selênio/metabolismo , Selênio/uso terapêutico , Raios XRESUMO
OBJECTIVE: Coxsackie B viruses (CVB3) are considered to be the most common etiologic agents of viral myocarditis. There are not any special anti-CVB3 drugs yet. From previous studies, the anti-CVB3 affect of sophora flavescens ait (SFA) had been discovered. Our experiment was to study the anti-CVB3 ability of SFA to cultured heating myocardial cells of CVB3 infected newborn rat. METHODS: The myocardial cells were divided into four groups: 1. infected group (n = 8), infected only with CVB3, not adding SFA; 2. treated group (n =8), infected with CVB3, adding SFA (100 microg/ml); 3. drug group (n=6), adding SFA (100 microg/ml) only; 4. control group (n = 6), not infected with CVB3, not adding SFA. RESULTS: The myocardial cells of the infected group had cell pathogenic effect (CPE) on the second day after virus inoculation, the CPE progressed rapidly from + to ++++. In contrast, no CPE in the other three groups was found. The LDH and SGOT of the infected group were higher than that in the other three groups, showing a significant difference (P <0.05). The virus titer of the infected group was higher than that of the treated group. There was no influence on normal myocardial cells if the concentration of SFA was lower than 300 microg/ml. When the concentration of SFA was 6.25 microg/ml 200 microg/ml, it showed protective effect on infected myocardial cells. CONCLUSIONS: The results of the experiment suggest that SFA might inhibit CVB3 replication in myocardial cells.