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Objective To explore the diagnosis and clinical characteristics of atypical severe pneumonia caused by Chlamydia abortus(C.abortus).Methods Clinical data of 4 patients diagnosed with atypical severe pneumonia caused by C.abortus in a hospital from January 2021 to November 2022 were collected.Clinical characteristics,dia-gnosis and treatment,and precautions of the disease were comprehensively analyzed.Results All 4 patients were male,aged 63-73 years old,with acute onset,high fever,cough and expectoration.Three patients had a history of contact with poultry,one patient had a history of contact with abortion goat.The interval between the emerging of clinical symptoms and the onset of acute respiratory failure in 4 patients was 1-6 days,and the oxygenation index(PaO2/FiO2)at admission was less than 200 mmHg,which gradually decreased with the progression of the disease,active support with a ventilator was necessary.Two patients had an increase in white blood cell count,4 had an in-crease in neutrophil percentage,3 had a mild decrease in platelet count.Among 4 patients,2,2,3 and 4 patients showed elevated levels of aspartate aminotransferase,alanine aminotransferase,creatine kinase,and serum creati-nine respectively,2 patients had mild hyponatremia,4 patients showed significant increase in C-reactive protein,procalcitonin,and interleukin-6 levels.Four patients'chest CT findings showed main involvement of single or mul-tiple lung lobes,with exudation and consolidation,and later involvement of multiple lobes of lung.The metageno-mic next-generation sequencing of bronchoalveolar lavage fluid detected the DNA sequence of C.abortus.Based on the clinical manifestations,contact history,chest CT,and metagenomic next-generation sequencing results of 4 pa-tients,the diagnosis was C.abortus.atypical severe pneumonia.After timely adjustment of the treatment of anti-in-fection regimen based on doxycycline,the patients'condition improved and were discharged.Conclusion C.abor-tus may also cause human pneumonia,which can lead to serious clinical outcome after infection.Patient had a histo-ry of animal contact should be alert to such diseases.Metagenomic next-generation sequencing can detect C.abortus.
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Many factors are reported to be involved in regulating the immunopathogenesis of schistosome infection. CD4+ T cell is one of the key players in the regulation of the liver granuloma formation by differentiation into different effector subsets including T helper (Th) 1, Th2, Th17, and T regulatory cells (Treg cells). Treg cells play an important suppressive role in immunopathology control and favor the pathogen to escape from the host immune assault. The functional activity of Tregs has been related to some autoimmune diseases including asthma and inflammatory bowel disease, which suggests that the manipulation of Tregs to restore their numbers and function may be therapeutic. However, interleukin-17 (IL-17) is a pro-inflammatory cytokine involved in the pathogenesis of many inflammatory and infectious conditions, including schistosomiasis. Therefore, a deeper understanding of the mechanisms of these immune regulations is necessary for the better control of pathology in schistosomiasis. In this paper, we review the Treg/Th17 balance and the immunology of schistosome infection.
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Esquistosomiasis , Linfocitos T Reguladores , Células Th17 , Animales , Linfocitos T CD4-Positivos/inmunología , Humanos , Schistosoma/inmunología , Esquistosomiasis/inmunología , Linfocitos T Reguladores/inmunología , Células Th17/inmunologíaRESUMEN
Many factors are reported to be involved in regulating the immunopathogenesis of schistosome infection. CD4+ T cell is one of the key players in the regulation of the liver granuloma formation by differentiation into different effector subsets including T helper (Th) 1, Th2, Th17, and T regulatory cells (Treg cells). Treg cells play an important suppressive role in immunopathology control and favor the pathogen to escape from the host immune assault. The functional activity of Tregs has been related to some autoimmune diseases including asthma and inflammatory bowel disease, which suggests that the manipulation of Tregs to restore their numbers and function may be therapeutic. However, interleukin-17 (IL-17) is a pro-inflammatory cytokine involved in the pathogenesis of many inflammatory and infectious conditions, including schistosomiasis. Therefore, a deeper understanding of the mechanisms of these immune regulations is necessary for the better control of pathology in schistosomiasis. In this paper, we review the Treg/Th17 balance and the immunology of schistosome infection.
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Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 μmol·L, SPD), alpha-difluoromethylornithine (2.5 mmol·L, DFMO), 4-Aminopyridine (40 μmol·L, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca ([Ca]) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca] and accelerated migration of IEC-6 cells, compared with the controls (P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca], but also restored IEC-6 cell migration to control level (P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased (P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.
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Animales , Ratas , Astragalus propinquus , Química , Atractylodes , Química , Línea Celular , Movimiento Celular , Medicamentos Herbarios Chinos , Química , Farmacología , Células Epiteliales , Biología Celular , Metabolismo , Intestinos , Biología Celular , Genética , Metabolismo , Poliaminas , Metabolismo , Polisacáridos , Química , Farmacología , Rizoma , Química , Transducción de Señal , Proteína de Unión al GTP rhoA , MetabolismoRESUMEN
Many factors are reported to be involved in regulating the immunopathogenesis of schistosome infection. CD4+ T cell is one of the key players in the regulation of the liver granuloma formation by differentiation into different effector subsets including T helper (Th) 1, Th2, Th17, and T regulatory cells (Treg cells). Treg cells play an important suppressive role in immunopathology control and favor the pathogen to escape from the host immune assault. The functional activity of Tregs has been related to some autoimmune diseases including asthma and inflammatory bowel disease, which suggests that the manipulation of Tregs to restore their numbers and function may be therapeutic. However, interleukin-17 (IL-17) is a pro-inflammatory cytokine involved in the pathogenesis of many inflammatory and infectious conditions, including schistosomiasis. Therefore, a deeper understanding of the mechanisms of these immune regulations is necessary for the better control of pathology in schistosomiasis. In this paper, we review the Treg/Th17 balance and the immunology of schistosome infection.
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Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 μmol·L, SPD), alpha-difluoromethylornithine (2.5 mmol·L, DFMO), 4-Aminopyridine (40 μmol·L, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca ([Ca]) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca] and accelerated migration of IEC-6 cells, compared with the controls (P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca], but also restored IEC-6 cell migration to control level (P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased (P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.
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Animales , Ratas , Astragalus propinquus , Química , Atractylodes , Química , Línea Celular , Movimiento Celular , Medicamentos Herbarios Chinos , Química , Farmacología , Células Epiteliales , Biología Celular , Metabolismo , Intestinos , Biología Celular , Genética , Metabolismo , Poliaminas , Metabolismo , Polisacáridos , Química , Farmacología , Rizoma , Química , Transducción de Señal , Proteína de Unión al GTP rhoA , MetabolismoRESUMEN
Aim To observe the effect of epidermal growth factor (EGF) on cell growth under different culture conditions using the real-time cell analyzer and EGF as a tool medicine to promote cell growth,and to provide reference for establishing pharmacokinetic model of IEC-6 cell growth (proliferation).Methods IEC-6 cell was inoculated on E-Plate 16 plate at a density of 1 × 104 cell/well and cultured in DMEM with 10% serum for 24 h,then replaced by serum-free DMEM culture (serum starvation)for 20 h,then the effects of different culture conditions on cell growth as well as EGF efficacy were observed.Results ① When the serum concentration was 10%,the cell growth index of EGF group(1,10,100 μg · L-1) after drug administration 24 h,48 h and 72 h was P > 0.05 compared with the blank group,suggesting that 10% serum culture could not reflect the efficacy of EGF.② When the serum concentration was 0%,EGF (1,10 μg · L-1) improved cell growth inhibition caused by serum-free cultivation,but could not recover it to normal level (the EGF group after drug administration 24 h,48 h and 72 h was P <0.01 compared with 5% serum),which suggested that serum-free culture could not reflect the EGF efficacy.③ 0%,0.5%,1% serum had different effects on cell growth,of which 0.5% serum could neither have obvious inhibition on cell growth,nor reflect the EGF effect due to promoting cell growth for a long time.④When the serum concentration was 0.5 %,the cell growth index of EGF groups after drug administration 24h,48h and 72h was P <0.01 compared with the blank group,suggesting that 0.5% serum culture could better reflect EGF efficacy.⑤The efficacy of EGF (10 μg· L-1) in promoting cell growth was confirmed by repeated validation of 0.5 % serum.Condusions A reference scheme of the IEC-6 cell growth (proliferative) pharmacological experimental model is established in the real-time cell analyzer:cells are cultured in DMEM with 10% serum for 24h,then in serum-free DMEM (serum starvation) for 20h,then after the adding of reagent,cells are cultured in DMEM with 0.5% serum for 48-72 h to observe its effect on cell growth (proliferation).
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We sought to investigate the underlying mechanism of action of the long noncoding RNA (lncRNA) LOC283070 in the development of androgen independence in prostate cancer. The interactions between LOC283070 and target proteins were investigated by RNA pull-down and RNA-binding protein immunoprecipitation (RIP) assays. Subcellular fractionation and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were used to detect the subcellular localization of LOC283070. Western blotting was performed to detect the expression of prohibitin 2 (PHB2). Luciferase activity assays were performed to evaluate the effects of LOC283070 and PHB2 on the androgen receptor (AR) signaling pathway. A methyl thiazolyl tetrazolium (MTT) assay and a growth curve assay were used to test cell viability. Flow cytometry was performed to analyze cell cycles. A transwell assay was employed to test cell migration. We identified PHB2 as an interaction partner of LOC283070 in the pull-down and RIP experiments. Furthermore, we confirmed that the enrichment of LOC283070 with PHB2 in androgen-independent LNCaP (LNCaP-AI) cells was much greater than that in LNCaP cells. Moreover, the expression of PHB2 was not significantly different between the two cell lines, and the expression of LOC283070 in the nuclei of the LNCaP-AI cells was significantly greater than that in the LNCaP cells. In vitro data revealed that PHB2 overexpression significantly inhibited AR activity and cell proliferation and migration and induced accumulation of prostate cancer cells in G0/G1 phase. Moreover, the overexpression of LOC283070 fully abrogated the effects of PHB2 overexpression. In conclusion, we found that LOC283070 can bind to PHB2 located in the nucleus and inhibit its effect, and this is one of the mechanisms by which LOC283070 is involved in the transition of LNCaP cells into androgen-independent cells.
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Humanos , Masculino , Andrógenos/metabolismo , Línea Celular Tumoral , Proliferación Celular/fisiología , Regulación Neoplásica de la Expresión Génica , Prohibitinas , Neoplasias de la Próstata/metabolismo , ARN Largo no Codificante/metabolismo , Receptores Androgénicos/metabolismo , Proteínas Represoras/metabolismo , Transducción de Señal/fisiologíaRESUMEN
BACKGROUND: Gender differences in the function and anatomical features of salivary glands are well known. However, specific gender differences in the biochemical composition and salivary flow rate (SFR) remain uncertain. Collection methods affect the assessment of the salivary composition and SFR, which are also highly affected by acid stimulation. METHODS: In the present study, we analyzed the differences in salivary characteristics of SFR, pH and salivary α-amylase (sAA) for 28 females and 27 males before and after citric acid stimulation, as measured by 3 different collection methods sequentially. RESULTS: Salivary pH values were significantly lower in females than that in males, both before and after stimulation, irrespective of collection methods. Salivary pH consistently increased after acid stimulation in both genders. Mean SFR in females before acid stimulation was significantly lower than that in males in all 3 samples collected. No gender difference in sAA was evident. CONCLUSION: Substantial gender differences in biochemistry and flow of saliva exist, and these findings are robust, as evidenced by reasonable consistency of the data among different saliva sampling methods.
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Ácido Cítrico/farmacología , Saliva/química , Adulto , Femenino , Humanos , Concentración de Iones de Hidrógeno , Masculino , Saliva/efectos de los fármacos , alfa-Amilasas Salivales/análisis , Factores Sexuales , Manejo de Especímenes/métodos , Manejo de Especímenes/normasRESUMEN
<p><b>OBJECTIVE</b>To compare the effect of citric acid stimulation on salivary alpha-amylase (sAA), total protein (TP), salivary flow rate, and pH value between Pi deficiency (PD) children and healthy children, thereby providing evidence for Pi controlling saliva theory.</p><p><b>METHODS</b>Twenty PD children were recruited, and 29 healthy children were also recruited at the same time. Saliva samples from all subjects were collected before and after citric acid stimulation. The sAA activity and amount, TP contents, salivary flow rate, and pH value were determined and compared.</p><p><b>RESULTS</b>(1) Citric acid stimulation was able to significantly increase salivary flow rate, pH value, sAA activities, sAA specific activity and sAA amount (including glycosylated and non-glycosylated sAA amount) in healthy children (P<0.05), while it could markedly increase salivary flow rate, pH value, and glycosylated sAA levels in PD children (P<0.05); (2) Although there was no statistical difference in determined salivary indices between the two groups (P>0.05), salivary indices except salivary flow rate and glycosylated sAA levels decreased more in PD children. There was statistical difference in sAA activity ratio, sAA specific activity ratio, and the ratio of glycosylated sAA levels between PD children and healthy children (P<0.05).</p><p><b>CONCLUSION</b>PD children had decreased response to citric acid stimulation.</p>
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Niño , Humanos , Ácido Cítrico , Usos Terapéuticos , Medicina Tradicional China , Saliva , alfa-Amilasas Salivales , Metabolismo , alfa-AmilasasRESUMEN
As the dilution procedure was applied, a simple, rapid and cost-effective high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for determination of aflatoxin B1, B2, G1, and G2 was successfully by performed in a total 83 samples of 10 traditional Chinese medicines (TCMs), which were collected from 5 different hospital pharmacies and 5 different medical stores in Guangzhou city. Matrix effects of these 10 TCMs were ranged from 80.23% to 115.5% in low, intermediate and high concentration levels, indicating that the negative effect was overcome in this study. Meanwhile, the analysis method was proved to be stable and reliable during the whole analysis using Semen Armeniacae Amarum spiked 3 concentration levels of standard solution as quality control samples and the RSD < 6.6% was obtained. The contamination levels of 83 investigated samples were 13.89% and 17.02% in hospital pharmacies and medical stores, respectively. The result was presented to provide relevant reference and supplement to those researchers in TCMs analysis and screening.
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Aflatoxina B1 , Aflatoxinas , Cromatografía Líquida de Alta Presión , Métodos , Contaminación de Medicamentos , Medicina Tradicional China , Control de Calidad , Espectrometría de Masas en Tándem , MétodosRESUMEN
The volatile components of roots and stems of Zanthoxylum nitidum were investigated by supercritical fluid carbon dioxide extraction (SFE-CO2) and gas chromatography-mass spectrometry(GC-MS). Thirty-one and fifty-one compounds were identified in the supercritical extracts from roots and stems of Z. nitidum, respectively, and total twenty-seven compounds were the common constituents. Among them, the major constituents in root and stem supercritical extracts were spathulenol (18.49 and 26.18%), n-hexadecanoic acid (14.24% and 12.79%), ar-tumerone (6.95% and 8.88%), oleic acid (8.39% and 5.71%) and hexanoic acid (4.39% and 7.78%). The in-vitro MTT assay showed that the volatile components of roots and stems of Z. nitidum did not exhibited any cytotoxic activity against human cancer Huh-7 and normal IEC-6 cells. These results indicated the same nature of the volatile constituents in the root and stem of Z. nitidum. This investigation may provide further evidence for expansion of medicinal parts of Z. nitidum.
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Animales , Humanos , Ratones , Línea Celular Tumoral , Supervivencia Celular , Cromatografía con Fluido Supercrítico , Medicamentos Herbarios Chinos , Química , Toxicidad , Cromatografía de Gases y Espectrometría de Masas , Métodos , Raíces de Plantas , Química , Tallos de la Planta , Química , Zanthoxylum , QuímicaRESUMEN
OBJECTIVE: To analyze the metabolic states of the lipids, protein, carbohydrate, and nucleic acid for chronic superficial gastritis patients of splenasthenic syndrome (SS), and to explore the pathogenesis mechanism of SS based on substance and energy metabolisms. METHODS: During June 2004 to March 2005, recruited were four chronic superficial gastritis patients of SS who visited at the First Hospital of Guangzhou University of Chinese Medicine and Guangdong Provincial Hospital of Traditional Chinese Medicine. Four healthy volunteers were recruited from Guangzhou University of Chinese Medicine. Their gastric mucosa was extracted to perform experiments of DNA microarray. The dual-channel DNA microarray data were mined and bioinformatics analyzed by BRB ArrayTools and IPA software. RESULTS: Fifteen genes were involved in substance and energy metabolisms in 20 differentially expressed genes, accounting for 75%.Among these genes, one gene was up-regulated, 14 genes down-regulated, and 11 genes were enzyme gene. Differentially expressed genes related to lipid metabolism included ACAA2 and CYP20A1, manifested as fatty acid catabolism and cholesterol transformation. Genes related to protein metabolism included ALDH9A1, ASL, ASS1, PCY-OX1L, RPS28, UBE2D2, UBXN1, B3GNT1, GCNT1, and PPP1R3C, manifested as decreased amino acid metabolism that may affect the biologic processes such as autonomic nerve, urea cycle, etc., reduced protein synthesis, increased ubiquitination of fault fold proteins, and decreased post-translated modification of glycosylation and dephosphorylation. Genes related to carbohydrate metabolism included PPP1R3C, B3GNT1, and GCNT1, manifested as decreased glycogen and glycan syntheses. Genes related to nucleic acid metabolism included RMI1, SMARCD3, and PARP1, manifested as degraded DNA duplication and transcription, and increased DNA damage repair. CONCLUSIONS: The metabolisms of the lipids, protein, carbohydrate, and nucleic acid in chronic superficial gastritis patients of SS obviously decreased, manifested mainly as down-regulated enzyme gene expression. We inferred that these might be one of the vital pathogenesis mechanisms for nutrition dysmetabolism of SS.
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Metabolismo Energético/genética , Gastritis/metabolismo , Perfilación de la Expresión Génica , Adulto , Estudios de Casos y Controles , Femenino , Gastritis/diagnóstico , Gastritis/genética , Expresión Génica , Humanos , Masculino , Medicina Tradicional China/métodos , Análisis de Secuencia por Matrices de Oligonucleótidos , Adulto JovenRESUMEN
<p><b>OBJECTIVE</b>To observe the effect of Sijunzi Decoction (SD) on the intestinal absorption of glucose in model rats of Pi-qi deficiency syndrome (PQDS).</p><p><b>METHODS</b>PQDS rat model was established by subcutaneous injection of reserpine from the neck. The body weight and urine D-xylose excretion rates were measured. The glucose uptake rate was measured by jejunum perfusion. The intestinal mucosa was collected. The glucose transporter-2 (GLUT2) protein and mRNA expression levels were detected.</p><p><b>RESULTS</b>Compared with the normal control group, the body weight and D-xylose excretion rates decreased in the model group. Meanwhile, the glucose uptake and GLUT2 protein and mRNA expression levels decreased in the model group. The aforesaid indices were improved in the SD group.</p><p><b>CONCLUSION</b>SD could promote the recovery of glucose uptake in the small intestine of reserpine induced PQDS rats.</p>
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Animales , Femenino , Ratas , Medicamentos Herbarios Chinos , Farmacología , Glucosa , Metabolismo , Transportador de Glucosa de Tipo 2 , Metabolismo , Absorción Intestinal , Mucosa Intestinal , Metabolismo , Intestino Delgado , Metabolismo , Qi , Ratas Sprague-Dawley , ReserpinaRESUMEN
<p><b>OBJECTIVE</b>To evaluate fungal contamination on the surface of Chinese herbal medicines and explore an appropriate method for fast and efficient identification of contaminant fungi.</p><p><b>METHOD</b>Chinese herbal medicines were first washed and the washing solution was plated onto potato dextrose agar (PDA) to obtain the pure isolates. For molecular identification, two new pairs of specific primers were designed according to ITS region of fungi genome sequences. The strains were identified through polymerase chain reaction (PCR) and sequence analysis.</p><p><b>RESULT</b>Fifty fungal strains were obtained from the surface of 15 Chinese herbal medicines with the percent of contaminated samples of 93.3%. Twenty-seven strains among them were successfully identified.</p><p><b>CONCLUSION</b>Fungal contamination on the surface of Chinese herbal medicines is quite common. Although different fungal species were isolated, the genus Aspergillus was the predominant. The primer pairs developed in this study are compatible and can be used to identify fungal species from the surface of Chinese herbal medicines.</p>
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Contaminación de Medicamentos , Medicamentos Herbarios Chinos , Hongos , Genética , Reacción en Cadena de la PolimerasaRESUMEN
A simple, rapid and cost-effective high-performance liquid chromatography-tandem mass spectrometry (LC-MS/ MS) method was established for simultaneous determination of aflatoxins (AFB1, AFB2, AFG1, AFG2) in Armeniacae Semen Amarum and the application was performance in 11 samples collected from different markets, medical stores and hospitals. The sample was extracted with 84% acetonitrile/water and 250 microL extraction was directly injected into a LC-MS/MS system without further purification procedure after being redissolved with methanol. The LC separation was performed on a C18 column with a linear gradient elution program of 4 mmol x L(-1) NH4 Ac-0.1% formic acid solution and menthol as the mobile phase. Selected reaction monitoring (SRM) was used for selective determination of the four aflatoxins on a triple quadruple mass spectrometer, which was operated in positive ionization modes. All the four aflatoxins showed a good linear relationship with r > 0.999 0, the average recoveries were between 87.88% and 102.9% and the matrix effect was ranged from 90.71% to 99.30% in low, intermediate and high levels. Furthermore, the higher recovery was obtained by the method reported in this study, comparing to the cleanup procedure with the Mycosep 226 purification column. Eleven samples collected were detected and the contamination levels of the AFB1 were between 1.590-2.340 microg x kg(-1) and the AF (B1 + B2 + G1 + G2) was ranged from 2.340 to 3.340 microg x kg(-1). In summary, the developed method was suitable to detect and screen AFB1, AFB2, AFG1, AFG2 in Armeniacae Semen Amarum.
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Aflatoxinas , Metabolismo , Cromatografía Líquida de Alta Presión , Métodos , Contaminación de Medicamentos , Hongos , Metabolismo , Prunus , Química , Microbiología , Semillas , Química , Microbiología , Espectrometría de Masas en Tándem , MétodosRESUMEN
<p><b>OBJECTIVE</b>To isolate and identify active neuraminidase constituents of Polygonum cuspidatum against influenza A (H1N1) influenza virus.</p><p><b>METHOD</b>On the basis of the bioassay-guided fractionation,such chromatographic methods as silica gel, sephadex LH-20 and HPLC were adopted to isolate active constituents of extracts from Polygonum cuspidatum, and their molecular structures were identifiied on the basis of their spectral data such as NMR and MS and physico-chemical properties.</p><p><b>RESULT</b>Seven compounds were isolated from the ethyl acetate extract of P. cuspidatum and identified as 2-methoxystypandrone (1), emodin (2), resveratrol (3), polydatin (4), emodin-8-O-beta-D-glucopyranoside (5), (E)-3, 5, 12-trihydroxystilbene-3-O-beta-D-glucopyranoside-2'-(3", 4", 5"-trihydroxybenzoate) (6) and catechin-3-O-gallate (7), respectively. Among them, the NA test showed that compounds 3, 6 and 7 had inhibitory effect against NAs activity, with IC50 values of 129.8, 44.8 and 21.3 micromol x L(-1), respectively. Moreover, the further CPE test showed compounds 6 and 7 had significant inhibitory effect against H1N influenza virus (EC50 = 5.9, 0.9 micromol x L(-1), respectively), with very low cytotoxicity to the host cells, their therapeutic selective index(SI) in MDCK cells ranged from 56 to 269.</p><p><b>CONCLUSION</b>The neuraminidase inhibitors against H1N1 anti-influenza virus isolated from extracts of P. cuspidatum on the basis of the bioassay-guided fractionation are significant in specifying their therapeutic material basis and drug R&D against influenza.</p>
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Humanos , Línea Celular , Medicamentos Herbarios Chinos , Química , Farmacología , Inhibidores Enzimáticos , Química , Farmacología , Fallopia japonica , Química , Subtipo H1N1 del Virus de la Influenza A , Gripe Humana , Virología , Estructura Molecular , NeuraminidasaRESUMEN
<p><b>OBJECTIVE</b>To analyze the metabolic levels of energy and substance in chronic superficial gastritis (CSG) patients of Pi deficiency syndrome (PDS) and of Pi-Wei hygropyrexia syndrome (PWHS), including lipid, protein, nucleic acid, carbohydrate, trace element, and energy metabolism, and to study the pathogenesis mechanism of PDS from substance and energy metabolisms.</p><p><b>METHODS</b>Recruited were 8 CSG patients who visited at First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine and Guangdong Provincial Hospital of Traditional Chinese Medicine from June 2004 to March 2005, including 4 patients of PDS and 4 of PWHS. Their gastric mucosae were used for experiments of DNA microarray. The dual-channel DNA microarray data were bioinformatically analyzed by BRB ArrayTools and IPA Software.</p><p><b>RESULTS</b>Obtained were fifty-six differentially expressed genes involved in substance and energy metabolisms with the expression fold more than 2, including 11 genes up-regulated and 45 genes down-regulated. Of them, genes correlated to lipid metabolism included CRLS1, LRP11, FUT9, GPCPD1, PIGL, SULT1A4, B3GNT1, ST8SIA4, and ACADVL, mainly involved in the metabolic processes of fatty acid, cholesterol, phospholipids, and glycolipid. Genes correlated to protein metabolism included ASRGL1, AARSD1, EBNA1BP2, PUM2, MRPL52, C120RF65, PSMB8, PSME2, UBA7, RNF11, FBXO44, ZFYVE26, CHMP2A, SSR4, SNX4, RAB3B, RABL2A, GOLGA2, KDELR1, PHPT1, ACPP, PTPRF, CRKL, HDAC7, ADPRHL2, B3GNT1, ST8SIA4, DDOST, and FUT9, mainly involved in the biosynthesis processes of protein, ubiquitination, targeted transport and post-translation modification. Genes correlated to nucleic acid metabolism included DFFB, FLJ35220, TOP2A, SF3A3, CREB3, CRTC2, NR1D2, MED6, GTF2IRD1, C1ORF83, ZNF773, and ZMYND11, mainly involved in DNA replication and repair, transcription regulation. Genes correlated to carbohydrate metabolism included AGL, B3GNT1, FUT9, ST8SIA4, SULT1A4, DDOST, and PIGL, mainly involved in glucogen degradation and glycoconjugate biosynthesis. Genes correlated to trace element metabolism included COMMD1, SLC39A6, FTL, CHRFAM7A, SCGN, and S100A6, mainly involved in ion metabolisms of copper, zinc, ferri, and calcium. Genes correlated to energy metabolism included AK3 and COX7B, mainly involved in mitochondria structure and oxidative phosphorylation processes.</p><p><b>CONCLUSION</b>The metabolic levels of energy and substance including lipid, protein, nucleic acid, carbohydrate, and trace element were obviously reduced in patients of PDS, which might be an important pathogenesis mechanism for its occurrence.</p>
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Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Metabolismo Energético , Genética , Gastritis , Diagnóstico , Genética , Metabolismo , Regulación de la Expresión Génica , Medicina Tradicional China , Análisis de Secuencia por Matrices de Oligonucleótidos , TranscriptomaRESUMEN
<p><b>OBJECTIVE</b>To study differential expression profiles of ribosomal protein (RP) genes in healthy subjects and ulcerative colitis (UC) patients of Pi-asthenic syndrome (PAS) and of dampness-heat syndrome (DHS), thus providing experimental bases for " Pi as the source of qi and blood" theory from the view of protein synthesis.</p><p><b>METHODS</b>RP genes arrays were made. The mucous membrane of colon was detected in four UC patients of PAS (UC-PAS), four UC patients of DHS (UC-DHS), and four healthy subjects (N), and data analyzed using BRB-TOOL Software Package (3.9). Bioinformatics analyses were conducted in differential genes.</p><p><b>RESULTS</b>Low-density RP gene chips were successfully produced, including 77 RP genes and two RP like genes (RPL26-like1 and RPL7-like1). There were twelve differential genes between UC (PAS+DHS) and N, all of which were down-regulated genes. There were nineteen differential genes between UC-DHS and N, all of which showed down-regulating tendency. There were three differential genes between UC-PAS and N, all of which were down-regulated genes. There were six differential genes between UC-PAS and UC-DHS, all of which were up-regulated genes. Cluster analysis showed that normal and UC samples of this chip can be classified according to gene expression profiles, and UC-PAS and UC-DHS can be classified by clustering. Various differential genes had a common transcription regulatory factor.</p><p><b>CONCLUSIONS</b>RP genes arrays were successfully produced. RP gene expressions were down-regulated in UC-PAS and UC-DHS. Corresponding gene expression profiles were shown in N, UC-PAS and UC-DHS.</p>
Asunto(s)
Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Estudios de Casos y Controles , Colitis Ulcerosa , Diagnóstico , Genética , Metabolismo , Perfilación de la Expresión Génica , Mucosa Intestinal , Metabolismo , Medicina Tradicional China , Análisis de Secuencia por Matrices de Oligonucleótidos , Proteínas Ribosómicas , Genética , Metabolismo , TranscriptomaRESUMEN
<p><b>OBJECTIVE</b>To determine the bioinformatical characteristics of differential gene expression in patients with chronic superficial gastritis (CSG) with the Pi-deficiency syndrome (PDS) and those of the non-Pi-deficiency syndrome (non-PDS), i.e. patients of CSG with Pi-Wei dampnese-heat syndrome and healthy persons.</p><p><b>METHODS</b>With the BRB-Array Tools software package, original data collection and bioinformatic: analysis of gene arrays were conducted in 6 CSG patients of PDS (CSG-PDS), 6 CSG patients of non-PDS (CSG-nPDS), and 6 healthy volunteers (Normal).</p><p><b>RESULTS</b>Compared with non-PDS, the gene expressions: in PDS with regards to protein synthesis, energy metabolism, immune reaction and ionic transport tended to be down-regulated, while those concerning secretion, cytoskeleton and ubiquitinization were up-regulated dominantly.</p><p><b>CONCLUSIONS</b>The two kinds of samples, CSG-PDS/Normal and CSG-PDS/CSG-nPDS, have their respective gene expression profiles with different characteristics. Gene expression profile has certain referential significance in syndrome classification.</p>