RESUMO
We reported a 28-year-old male patient who had been admitted to a local hospital for several times in the past four years because of recurrent fever and cough. Each chest CT scan during hospitalization showed consolidation accompanied by exudation and mild pleural effusion. After treatment, the consolidation apparently absorbed, but similar symptoms recurred within half a year, and the new consolidation appeared. For this reason, he was diagnosed with tuberculosis or bacterial pneumonia several times in other hospitals, and was hospitalized two to three times a year. Finally, he was diagnosed with chronic granulomatous disease (CGD) with CYBB gene mutation through whole-exome sequencing.
Assuntos
Doença Granulomatosa Crônica , Derrame Pleural , Masculino , Humanos , Adulto , Doença Granulomatosa Crônica/genética , Doença Granulomatosa Crônica/diagnóstico , NADPH Oxidase 2/genética , MutaçãoRESUMO
Hidradenitis suppurativa (HS) is a chronic disease of follicular occlusion. It involves the axilla, groin, perianal and perineal regions, and is characterized by recurrent draining sinuses, skin abscesses and disfiguring scars. Loss-of-function mutations in the genes encoding γ-secretase have been identified as a cause of HS. We collected skin samples from three patients with HS from a Chinese family carrying a NCSTN mutation (c.1258C>T (p.Q420X)) and three unrelated healthy controls (HCs). Expression level of nicastrin in skin tissue and cultured keratinocytes and fibroblasts of patients and HCs was determined by real-time quantitative PCR and western blotting. We found that the mRNA and protein levels of nicastrin were significantly reduced in the whole skin, epidermis, dermis, and cultured keratinocytes and fibroblasts compared with HCs. Therefore, we conclude that haploinsufficiency of the NCSTN gene caused by the nonsense mutation c.1258C>T (p.Q420X) contributes to the occurrence of HS in this family.
Assuntos
Secretases da Proteína Precursora do Amiloide/genética , Códon sem Sentido , Haploinsuficiência/genética , Hidradenite Supurativa/genética , Glicoproteínas de Membrana/genética , Povo Asiático/genética , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Previously, from the human intestinal flora we isolated the bacterial strain Bacteroides uniformis ZL1, which could convert secoisolariciresinol diglucoside (SDG) to its aglycone secoisolariciresinol (SECO) in vivo. In this study, 24 putative ß-glucosidase genes were screened from the genome of B. uniformis ATCC 8492, which were used as templates to design PCR primers for the target genes in B. uniformis ZL1. Fifteen genes (bgl1-bgl15) were amplified from strain ZL1, and among them we identified bgl8 as the gene encoding the SDG-hydrolyzing ß-glucosidase. We sequenced the bgl8 gene, cloned it into the expression vector and then transformed Escherichia coli to construct the recombinant bacteria that could synthesize the target ß-glucosidase (BuBGL8). We purified and characterized BuBGL8, which showed maximal activity and stability under the culture conditions of pH 6.0 and 30 °C. SDG (2.0 mg/ml) was converted to SECO by both the purified BuBGL8 (0.035 mg/ml) and crude enzyme extract (0.23 mg crude protein/ml) with the efficiency of more than 90 % after 90 min at the reaction conditions. This is, to our knowledge, the first report of using recombinant bacteria to synthesize the SDG-hydrolyzing ß-glucosidase, which could be used to produce SECO from SDG conveniently and highly efficiently.
Assuntos
Bacteroides/enzimologia , Butileno Glicóis/metabolismo , Glucosídeos/metabolismo , Lignanas/metabolismo , beta-Glucosidase/genética , beta-Glucosidase/metabolismo , Sequência de Aminoácidos , Bacteroides/genética , Clonagem Molecular , DNA Bacteriano/química , DNA Bacteriano/genética , Estabilidade Enzimática , Escherichia coli/genética , Expressão Gênica , Concentração de Íons de Hidrogênio , Hidrólise , Modelos Moleculares , Simulação de Acoplamento Molecular , Dados de Sequência Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , TemperaturaRESUMO
AIMS: Secoisolariciresinol (SECO) is increasingly recognized for potential clinical application because of its preventive effects against breast and colon cancers, atherosclerosis and diabetes, and its production through biotransformation has been attempted. However, previously reported bacteria all required stringent anaerobic culture conditions, precluding large-scale production. Here, we report the isolation and characterization of bacteria that produce SECO under less stringent anaerobic culture conditions. METHODS AND RESULTS: Using defatted flaxseed as raw material, we isolated a facultative anaerobic bacterium from human faeces that hydrolysed secoisolariciresinol diglucoside-3-hydroxy-3-methyl glutaric acid (SDG-HMGA) oligomers in flaxseed to produce SECO. Both conventional assays and 16S rRNA gene sequence analysis demonstrated its close relatedness with Bacteroides uniformis. The transformation efficiency of SDG in defatted flaxseed to SECO was more than 80% by this bacterial strain. We investigated factors that might influence fermentation, such as redox potential and pH, for large-scale fermentation of defatted flaxseed to produce SECO. CONCLUSIONS: The method to produce SECO through biotransformation of defatted flaxseed with this bacterial strain is highly efficient and economic. SIGNIFICANCE AND IMPACT OF THE STUDY: This bacterial strain can transform SDG to SECO under less stringent anaerobic culture conditions, which will greatly facilitate industry-scale production of SECO.
Assuntos
Bacteroidaceae/metabolismo , Butileno Glicóis/metabolismo , Fermentação , Linho/química , Lignanas/metabolismo , Adulto , Bacteroidaceae/genética , Bacteroidaceae/isolamento & purificação , Biotransformação , Butileno Glicóis/química , Butileno Glicóis/isolamento & purificação , Cromatografia Líquida de Alta Pressão , Fezes/microbiologia , Feminino , Glucosídeos/metabolismo , Humanos , Hidrólise , Masculino , Filogenia , RNA Ribossômico 16S/genética , Adulto JovemRESUMO
BACKGROUND: Vascular endothelial growth factor (VEGF) promotes angiogenesis and plays important roles in neovascularization and development of tissues. VEGF receptors (VEGFRs) are high-affinity receptors for VEGF and are originally considered specific to endothelial cells. We have previously shown that keratinocytes from human normal skin express VEGFRs. This poses the question of whether these receptors are also expressed by epidermal appendages, as epidermal appendages are lined with epithelial cells. OBJECTIVE: To investigate the expression of VEGFR-2 compare with VEGF in epidermal appendages, including hair follicles, eccrine sweat glands and sebaceous glands. METHODS: Monoclonal antibodies to VEGF and VEGFR-2 were used for immunohistochemical examination of cryostat-cut sections of normal human skin specimens from 11 donors undergoing cosmetic surgery. RESULTS: Immunoreactivities for VEGF and VEGFR-2 principally showed parallel intense expression in anagen hair follicle (including outer root sheat, inner root sheath, dermal papillae epidermal matrix), sebaceous glands (ductal and secretory portions) and eccrine sweat glands (ductal and secretory portions), respectively. In particular, abundant expression of VEGF was found in the follicular basement membrane zone surrounding the bulb matrix and in the ductal and secretory portions of eccrine sweat glands. CONCLUSION: A potential VEGF/VEGFR-2 autocrine pathway may be defined by the coexpression of VEGF and VEGFR-2 in human skin epidermal appendages.
Assuntos
Epiderme/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Adolescente , Adulto , Idoso , Membrana Basal/metabolismo , Criança , Glândulas Écrinas/metabolismo , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Folículo Piloso/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Glândulas Sebáceas/metabolismo , Adulto JovemRESUMO
The novel 2DL4*00106 allele differs from the closest allele 2DL4*00102 by a silent mutation.
Assuntos
Alelos , Povo Asiático/genética , Etnicidade/genética , Receptores KIR2DL4/genética , Éxons/genética , Teste de Histocompatibilidade , HumanosRESUMO
The novel 2DL4*040 allele differs from the closest allele 2DL4*00102 by a missense mutation.
Assuntos
Alelos , Povo Asiático/genética , Etnicidade/genética , Receptores KIR2DL4/genética , Sequência de Bases , Éxons/genética , HumanosRESUMO
The novel KIR2DL4*039 allele differs from the closest allele 2DL4*00602 by a single non-synonymous mutation.
Assuntos
Alelos , Povo Asiático/genética , Etnicidade/genética , Receptores KIR2DL4/genética , Sequência de Bases , Éxons/genética , HumanosAssuntos
Alopecia/genética , Ictiose/genética , Metaloendopeptidases/genética , Fotofobia/genética , Adulto , Criança , Feminino , Humanos , Masculino , Linhagem , SíndromeRESUMO
Two novel phenanthrene derivatives, aristololactam IVa (1) and 9-hydroxy aristolochic acid I (2) were isolated from the stem and leaves of Anstolochia contorta Bunge, together with 17 known compounds (3-19). The structures of these compounds were determined by spectroscopic analysis. The phenanthrenes obtained were tested for cytotoxicity against renal proximal tubular epithelial cell line (HK-2). Aristololactam IVa and 7-methoxy aristololactam IV were found to have strong cytotoxic activity against HK-2 cells with a potency similar to or even stronger than those of aristolochic acid I and aristololactam I.
Assuntos
Aristolochia/química , Ácidos Aristolóquicos/química , Rim/patologia , Fenantrenos/química , Ácidos Aristolóquicos/isolamento & purificação , Linhagem Celular , Humanos , Túbulos Renais Proximais/citologia , Túbulos Renais Proximais/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Conformação Molecular , Fenantrenos/isolamento & purificação , Folhas de Planta/química , Caules de Planta/química , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Espectrofotometria Infravermelho , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Endothelin-1 (ET-1) acts as a key regulator of vasoconstriction and fibrosis. Many previous studies have focused on the role of ET-1 in scleroderma (systemic sclerosis, SSc). We investigated the effects of ET-1 on the production of extracellular matrix in SSc and normal skin fibroblasts. Primary cultured dermal fibroblasts from SSc patients and healthy controls were treated with ET-1 (25 ng/mL) for 0 min, 15 min, 1 h, 24 h, 48 h and 72 h, respectively. Our results showed that, in SSc fibroblasts, ET-1 upregulated collagen type I, connective tissue growth factor (CTGF), type I plasminogen activator inhibitor (PAI-1) and pAkt in a time-dependent manner within 72 h; in normal fibroblasts, 25 ng/mL ET-1 stimulation correlated with high levels of CTGF, PAI-1 and pAkt. The secretion of fibronectin (FN), collagen type I, and PAI-1 is markedly increased in the supernatant of both SSc fibroblasts and normal fibroblasts. Furthermore, ET-1 phosphorylates Smad2 and Smad3 in normal fibroblasts, but not in SSc fibroblasts. In conclusion, our results demonstrated that ET-1 may induce fibrosis in dermal fibroblasts through Akt signals.
Assuntos
Endotelina-1/farmacologia , Fibroblastos/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/genética , Escleroderma Sistêmico/genética , Pele/efeitos dos fármacos , Adulto , Estudos de Casos e Controles , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Fator de Crescimento do Tecido Conjuntivo/genética , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Endotelina-1/metabolismo , Feminino , Fibroblastos/metabolismo , Fibroblastos/patologia , Fibronectinas/biossíntese , Fibronectinas/metabolismo , Fibrose , Regulação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Fosforilação , Inibidor 1 de Ativador de Plasminogênio/genética , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Cultura Primária de Células , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Escleroderma Sistêmico/metabolismo , Escleroderma Sistêmico/patologia , Transdução de Sinais , Pele/metabolismo , Pele/patologia , Proteína Smad2/genética , Proteína Smad2/metabolismo , Proteína Smad3/genética , Proteína Smad3/metabolismoRESUMO
Three triterpenoids, 8alpha-hydroxyfernan-25,7beta-olide, 3alpha-hydroxy-4alpha-methoxyfilicane and 19alpha-hydroxyferna-7,9(11)-diene were isolated from the fresh fronds of Adiantum caudatum. Their structures were elucidated by spectroscopic techniques. Thirteen known triterpenoids were also identified from this fern.
Assuntos
Gleiquênias/química , Triterpenos/isolamento & purificação , Estrutura Molecular , Análise Espectral , Triterpenos/químicaRESUMO
Crude drugs derived from Adiantum species are used as febrifuge, antidote, diuretic, tonic, etc. Some commercial samples of these drugs are composed of finely cut ultimate pinnules only, which have false indusia and spores. In this paper, in order to establish a classification method based on the characteristics of the ultimate pinnules and their attachments, the morphological study using stereoscope and scanning electron microscope, and X-ray microanalysis using an electron probe microanalyzer were carried out on the false indusia and spores, and the ultimate pinnules, respectively, of 19 Adiantum species. The results showed that examined all species could be distinguished from each other by the following characteristics: in the false indusium, the shape and the presence or absence of hairs; in the spore, the shape, the ornamentation, the ratio of the laesura, and the equatorial diameter; in the X-ray images of the ultimate pinnule, the distributional patterns of silicon and calcium. The distributional patterns of silicon were due to the presence of spicular cells, hairs and papillae, and calcium was present as crystals of calcium oxalate. The average content of silicon in the ultimate pinnules of Adiantum species was 1.99%.
Assuntos
Cálcio/análise , Plantas Medicinais/classificação , Silício/análise , Esporos/ultraestrutura , Microanálise por Sonda Eletrônica , Microscopia Eletrônica de Varredura , Plantas Medicinais/química , Plantas Medicinais/fisiologiaRESUMO
In the previous paper, Tu-er-feng, one of Chinese folk medicines used locally in Sichuan prov., derived from the whole plants of Gerbera piloselloides of family Compositae, was pharmacognostically reported. In the recent markets, besides this known material, Tu-er-feng made of different components are found. In this paper, the Ainsliaea derivatives are studied to clarify the botanical origins; comparing anatomically with leaves and petioles of thirteen Ainsliaea species growing wildly in Sichuan prov. The result shows that A. glabra and A. rubrinervis are the ingredients.
Assuntos
Medicamentos de Ervas Chinesas , Plantas Medicinais/anatomia & histologiaRESUMO
Tu-er-feng is one of famous Chinese folk medicines in Sichuan prov. for common cold with cough, rheumatism, etc. Its sources are said to be either whole plants of some Gerbera or Ainsliaea species of family Compositae. In the recent markets, two types of Tu-er-feng are surely available. In this paper, Tu-er-feng derived from Gerbera species are studied to clarify the botanical origin; comparing mainly with the internal morphologies of the leaves and roots of G. piloselloides, G. delavayi, G. nivea, G. anandria (= Leibnitzia anandria) and G. jamesonii. As the result, G. piloselloides is determined as the botanical origin of Tu-er-feng obtained from the recent 16 markets.
Assuntos
Medicamentos de Ervas Chinesas , Plantas Medicinais/anatomia & histologiaRESUMO
OBJECTIVE: Based on an to investigation and identification of commercial samples to find out the original species and resources of Xue Lianhua. METHOD: Botanical and macroscopical identification. RESULT: The main original species of the plant are Saussurea involucrata, S. medusa, S. gossypiphora, S. polylada, S. laniceps, S. tridactyla, S. gnaphaloides, S. kingii, S. obvallata, S. tangutica, S. globosa, S. longifolia, Soroseris erysimoides, So. glomerata and So. umbrella. CONCLUSION: The flower and stem of S. involucrata and S. polylada and the whole herb of S. medusa and S. gossypiphora are used as Xue Lianhua in most areas of China.
Assuntos
Plantas Medicinais , Saussurea , Asteraceae/anatomia & histologia , Asteraceae/classificação , China , Conservação dos Recursos Naturais , Plantas Medicinais/anatomia & histologia , Plantas Medicinais/classificação , Saussurea/anatomia & histologia , Saussurea/classificaçãoRESUMO
OBJECTIVE: To study the chemical compositions of the root and stem of Patrinia scabiasaefolia. METHOD: Five compounds were obtained from the ethy acetate extract of the root and stem of P. scabiasaefolia by silica column chromatography, and identified by chemical and spectroscopic analysis. RESULT: The compounds were identified as 2'-acetyl-3-O-beta-arabinopyranosyl-hederagenin, 28-O-beta-D-glucopyranosyl-oleanolic acid, 3-O-alpha-L-arabinopyranosyl-(1 -->3)-beta-D-xylopyranosyl-hederagenin and 3-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl-oleanolic acid. CONCLUSION: These compounds were obtained from the plant of P. scabiasaefolia for the first time.
Assuntos
Patrinia/química , Plantas Medicinais/química , Saponinas/isolamento & purificação , Raízes de Plantas/química , Caules de Planta/química , Saponinas/químicaRESUMO
OBJECTIVE: To study the chemical constituents of the roots and rhizomes of Asarum longerhizomatosum. METHOD: Chromatography and spectral analysis were used to isolate the constituents and elucidate their structures. RESULT: Five compounds were isolated from the ethanol extracts of the roots and rhizomes, and identified as asarone(I), beta-sitosterol(II), 2,4,5-trimethoxybenzaldehyde(III), 4-(2,4,5-trimethoxyphenyl)-3-en-butylone(IV) and 3 beta-hydroxystigmast-5-en-7-one(V). CONCLUSION: All the compounds were isolated from the plant for the first time, and IV is a new natural product.
Assuntos
Anisóis/isolamento & purificação , Asarum/química , Plantas Medicinais/química , Derivados de Alilbenzenos , Anisóis/química , Raízes de Plantas/química , Rizoma/química , Sitosteroides/química , Sitosteroides/isolamento & purificaçãoRESUMO
BACKGROUND AND PURPOSE: The combination of Chinese herbs, Astragali Radix and Angelicae Sinensis Radix, could alleviate renal interstitial fibrosis. Astragaloside IV (AS-IV) and ferulic acid (FA) are the two major active constituents in this combination. In this study, we employed rats with unilateral ureteral obstruction to determine whether AS-IV and FA have the same renoprotective effects and investigated the mechanisms of this action. EXPERIMENTAL APPROACH: Renal pathological changes were evaluated after treatment with AS-IV, FA or AS-IV + FA (AF) for 10 days. Meanwhile, the expression of transforming growth factor ß(1) (TGF-ß(1) ), fibronectin, α-smooth muscle actin (α-SMA), phosphorylation of c-Jun NH(2) -terminal kinase (p-JNK) and nitric oxide (NO) production in kidney were determined. The expressions of fibronectin, α-SMA, mitogen-activated protein kinases [JNK, extracellular signal-regulated kinases (ERK), P38] in TGF-ß(1) -treated NRK-49F cells or interleukin-1-treated HK-2 cells after AS-IV, FA or AF were assessed. KEY RESULTS: AF alleviated the infiltration of mononuclear cells, tubular atrophy and interstitial fibrosis; reduced the expression of fibronectin, α-SMA, TGF-ß(1) and p-JNK; and dramatically increased the production of NO in obstructed kidneys. Neither AS-IV nor FA alone improved renal damage, but both increased NO production. AF inhibited α-SMA and fibronectin expression in NRK-49F or HK-2 cells. Furthermore, AF significantly inhibited IL-1ß-induced JNK phosphorylation, without affecting ERK or P38 phosphorylation. Neither AS-IV nor FA alone had any effect on the cells. CONCLUSIONS AND IMPLICATIONS: AS-IV synergizes with FA to alleviate renal tubulointerstitial fibrosis; this was associated with inhibition of tubular epithelial-mesenchymal transdifferentiation (EMT) and fibroblast activation, as well as an increase in NO production in the kidney.