Bioactive metabolites in of Ginkgo biloba leaves: variations by seasonal, meteorological and soil / Metabólitos bioativos em folhas de Ginkgo biloba: variações sazonais, meteorológicas e do solo

Abstract Ginkgo biloba is a traditional Chinese herbal medicine containing multiple components that contribute to its notable bioactivities. Variations of seasonal, meteorological and planting soil on the phytochemicals contents in G. biloba leaves due to the effects of growth meteorological and soil parameters were investigated in this study. The leaves of G. biloba were collected from different months and place in Zhejiang province, the contents of flavones (quercetin, kaempferol and isorhamnetin) and terpene lactones (bilobalide, ginkgolides A, B and C) were quantified by high performance liquid chromatography (HPLC) and the evaporative light scattering detector (ELSD) method. The established methods were validated with good linearity, precision, repeatability, stability, and recovery. Comprehensive analysis suggested the proper harvest time for G. biloba was in October of Zhejiang province. The result of correlation analysis with meteorological factors shows that the temperature and precipitation have non-significant effect on the main components of G. biloba. In addition, the type and content (Mn and Zn) of the soil showed significantly effect on the content of flavonoids and terpene lactones. This study enriched the knowledge on the development and utilization value of the G. biloba leaves and was useful for determining the optimal harvest time and growing condition.

Resumo Ginkgo biloba é um fitoterápico tradicional da medicina chinesa que contém vários componentes que contribuem para suas notáveis bioatividades. Variações sazonais, meteorológicos e de plantio do solo sobre os teores fitoquímicos em folhas de G. biloba, devido aos efeitos do crescimento meteorológico e parâmetros do solo, foram investigadas neste estudo. As folhas de G. biloba foram coletadas em diferentes meses e na província de Zhejiang, os teores de flavonas (quercetina, kaempferol e isorhamnetina) e lactonas terpênicas (bilobalida, ginkgolídeos A, B e C) foram quantificados por Cromatografia Líquida de Alta Eficiência (CLAE) e pelo método do detector de dispersão da luz por evaporação. Os métodos estabelecidos foram validados com boa linearidade, precisão, repetibilidade, estabilidade e recuperação. Uma análise abrangente sugeriu que o tempo de colheita adequado para G. biloba foi em outubro na província de Zhejiang. O resultado da análise de correlação com fatores meteorológicos mostra que a temperatura e a precipitação não têm efeito significativo sobre os principais componentes de G. biloba. Além disso, o tipo e composição (Mn e Zn) do solo apresentaram efeito significativo sobre o teor de flavonoides e lactonas terpênicas. Este estudo enriqueceu o conhecimento sobre o valor de desenvolvimento e utilização das folhas de G. biloba e foi útil para determinar o melhor tempo de colheita e condição de crescimento.

Bioactive metabolites in of Ginkgo biloba leaves: variations by seasonal, meteorological and soil

Abstract Ginkgo biloba is a traditional Chinese herbal medicine containing multiple components that contribute to its notable bioactivities. Variations of seasonal, meteorological and planting soil on the phytochemicals contents in G. biloba leaves due to the effects of growth meteorological and soil parameters were investigated in this study. The leaves of G. biloba were collected from different months and place in Zhejiang province, the contents of flavones (quercetin, kaempferol and isorhamnetin) and terpene lactones (bilobalide, ginkgolides A, B and C) were quantified by high performance liquid chromatography (HPLC) and the evaporative light scattering detector (ELSD) method. The established methods were validated with good linearity, precision, repeatability, stability, and recovery. Comprehensive analysis suggested the proper harvest time for G. biloba was in October of Zhejiang province. The result of correlation analysis with meteorological factors shows that the temperature and precipitation have non-significant effect on the main components of G. biloba. In addition, the type and content (Mn and Zn) of the soil showed significantly effect on the content of flavonoids and terpene lactones. This study enriched the knowledge on the development and utilization value of the G. biloba leaves and was useful for determining the optimal harvest time and growing condition.

Resumo Ginkgo biloba é um fitoterápico tradicional da medicina chinesa que contém vários componentes que contribuem para suas notáveis bioatividades. Variações sazonais, meteorológicos e de plantio do solo sobre os teores fitoquímicos em folhas de G. biloba, devido aos efeitos do crescimento meteorológico e parâmetros do solo, foram investigadas neste estudo. As folhas de G. biloba foram coletadas em diferentes meses e na província de Zhejiang, os teores de flavonas (quercetina, kaempferol e isorhamnetina) e lactonas terpênicas (bilobalida, ginkgolídeos A, B e C) foram quantificados por Cromatografia Líquida de Alta Eficiência (CLAE) e pelo método do detector de dispersão da luz por evaporação. Os métodos estabelecidos foram validados com boa linearidade, precisão, repetibilidade, estabilidade e recuperação. Uma análise abrangente sugeriu que o tempo de colheita adequado para G. biloba foi em outubro na província de Zhejiang. O resultado da análise de correlação com fatores meteorológicos mostra que a temperatura e a precipitação não têm efeito significativo sobre os principais componentes de G. biloba. Além disso, o tipo e composição (Mn e Zn) do solo apresentaram efeito significativo sobre o teor de flavonoides e lactonas terpênicas. Este estudo enriqueceu o conhecimento sobre o valor de desenvolvimento e utilização das folhas de G. biloba e foi útil para determinar o melhor tempo de colheita e condição de crescimento.

Histidine-tryptophan-ketoglutarate solution decreases mortality and morbidity in high-risk patients with severe pulmonary arterial hypertension associated with complex congenital heart disease: an 11-year experience from a single institution

Cardioplegic reperfusion during a long term ischemic period interrupts cardiac surgery and also increases cellular edema due to repeated solution administration. We reviewed the clinical experiences on myocardial protection of a single perfusion with histidine-tryptophan-ketoglutarate (HTK) for high-risk patients with severe pulmonary arterial hypertension associated with complex congenital heart disease. This retrospective study included 101 high-risk patients undergoing arterial switch operation between March 2001 and July 2012. We divided the cohort into two groups: HTK group, myocardial protection was carried out with one single perfusion with HTK solution; and St group, myocardial protection with conventional St. Thomas' crystalloid cardioplegic solution. The duration of cardiopulmonary bypass did not differ between the two groups. The mortality, morbidity, ICU stay, post-operative hospitalization time, and number of transfusions in HTK group were lower than those in St group (P<0.05). Univariate and multivariate analysis showed that HTK is a statistically significant independent predictor of decreased early mortality and morbidity (P<0.05). In conclusion, HTK solution seems to be an effective and safe alternative to St. Thomas' solution for cardioplegic reperfusion in high-risk patients with complex congenital heart disease.

Fibrilación auricular y su asociación con la declinación cognitiva, deterioro cognitivo y demencia en ausencia de accidente cerebrovascular clínico: revisión de la evidencia y posibles mecanismos / Atrial fibrillation and its association with cognitive decline, cognitive impairment, and dementia in the absence of clinical stroke: a review of the evidence and possible mechanisms

La fibrilación auricular (FA) es la arritmia más frecuente en la población adulta. Es sabido que la FA se asocia con un aumento del riesgo de presentar muchos eventos adversos, entre los que se incluyen accidente cerebrovascular, insuficiencia cardíaca, muerte por todas las causas, así como muerte súbita cardíaca. Más recientemente también ha habido un creciente interés en la asociación de FA con deterioro cognitivo y demencia. En esta revisión narrativa describimos la actual evidencia a favor de la relación entre la FA y la declinación cognitiva, el deterioro cognitivo o la demencia en pacientes sin antecedentes de accidente cerebrovascular. Aun cuando no queda muy claro cuáles son los mecanismos exactos que explican la asociación entre la FA y la declinación cognitiva, los mecanismos propuestos incluyen isquemia silente, estado pro inflamatorio, microsangrados cerebrales e hipoperfusión cerebral. Se necesitan más estudios para dilucidar los mecanismos de base que faciliten el descubrimiento de alguna estrategia de prevención.

Atrial fibrillation (AF) is the most common arrhythmia in the adult population. AF is widely known to be associated with an increased risk of many adverse outcomes including stroke, heart failure, all-cause death, as well as sudden cardiac death. More recently, there has also been growing interest in the association of AF with cognitive impairment and dementia. In this narrative review, we describe the current evidence that supports the relationship of AF to cognitive decline, cognitive impairment, or dementia in patients without a history of stroke. The exact mechanisms underlying the association of AF with cognitive decline are unclear; however, proposed mechanisms include silent ischemia, pro-inflammatory state, cerebral microbleeds, and cerebral hypoperfusion. Further research is needed to elucidate the underlying mechanisms to facilitate discovery of prevention strategies.

Modulatory effects of taurine on jejunal contractility

Taurine (2-aminoethanesulfonic acid) is widely distributed in animal tissues and has diverse pharmacological effects. However, the role of taurine in modulating smooth muscle contractility is still controversial. We propose that taurine (5-80 mM) can exert bidirectional modulation on the contractility of isolated rat jejunal segments. Different low and high contractile states were induced in isolated jejunal segments of rats to observe the effects of taurine and the associated mechanisms. Taurine induced stimulatory effects on the contractility of isolated rat jejunal segments at 3 different low contractile states, and inhibitory effects at 3 different high contractile states. Bidirectional modulation was not observed in the presence of verapamil or tetrodotoxin, suggesting that taurine-induced bidirectional modulation is Ca2+ dependent and requires the presence of the enteric nervous system. The stimulatory effects of taurine on the contractility of isolated jejunal segments was blocked by atropine but not by diphenhydramine or by cimetidine, suggesting that muscarinic-linked activation was involved in the stimulatory effects when isolated jejunal segments were in a low contractile state. The inhibitory effects of taurine on the contractility of isolated jejunal segments were blocked by propranolol and L-NG-nitroarginine but not by phentolamine, suggesting that adrenergic β receptors and a nitric oxide relaxing mechanism were involved when isolated jejunal segments were in high contractile states. No bidirectional effects of taurine on myosin phosphorylation were observed. The contractile states of jejunal segments determine taurine-induced stimulatory or inhibitory effects, which are associated with muscarinic receptors and adrenergic β receptors, and a nitric oxide associated relaxing mechanism.

Nontyphoidal Salmonella urinary tract infection in a case of hyperparathyroidism and nephrocalcinosis / Infección de las vías urinaria por Salmonella no tifoidea en un caso de hiperparatiroidismo y nefrocalcinosis

Nontyphoidal Salmonella infections often present with self-limited gastroenteritis. Extraintestinal focal infections are uncommon but have high mortality and morbidity. Urinary tract infection caused by nontyphoidal Salmonella is usually associated with structural abnormalities of the urinary tract. Nephrocalcinosis and nephrolithiasis are the major risk factors. Although primary hyperparathyroidism has been reported to increase the risk of nephrocalcinosis and nephrolithiasis, little is known about the association between hyperparathyroidism and Salmonella urinary tract infection. We report the case of a 37-year old man who had a history of primary hyperparathyroidism and bilateral nephrocalcinosis and who developed urinary tract infection. Salmonella Group D was isolated from his urine specimen. Salmonella should be considered as a possible causality organism in patients with primary hyperparathyroidism and nephrocalcinosis who develop urinary tract infection. These patients need to be aware of the potential risks associated with salmonellosis.

Las infecciones por Salmonella no tifoidea se presentan a menudo con gastroenteritis auto-limitada. Las infecciones extra-intestinales focales son poco frecuentes, pero tienen una alta mortalidad y morbilidad. La infección de las vías urinarias causada por la Salmonella no tifoidea se asocia generalmente a anomalías estructurales de las vías urinarias. La nefrocalcinosis y la nefrolitiasis son los principales factores de riesgo. Aunque se ha reportado que el hiperparatiroidismo primario aumenta el riesgo de la nefrocalcinosis y la nefrolitiasis, poco se sabe sobre la asociación entre el hiperparatiroidismo y la infección de las vías urinarias por Salmonella. Damos a conocer aquí el caso de un hombre de 37 años con una historia de hiperparatiroidismo primario y nefrocalcinosis bilateral, que desarrolló una infección de las vías urinarias. La Salmonella del grupo D fue aislada de su muestra de orina. La Salmonella se debe considerar como un posible organismo de causalidad en pacientes con hiperparatiroidismo primario y nefrocalcinosis que desarrollan infección del tracto urinario. Estos pacientes necesitan tomar conciencia de los riesgos potenciales asociados con la salmonellosis.

The surface domain of neisserial IgA1 protease plays a role in induction of bacterial adherence to human epithelial cells

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Cloning, sequence analysis, and expression of cDNA coding for the major house dust mite allergen, Der f 1, in Escherichia coli

Our objective was to clone, express and characterize adult Dermatophagoides farinae group 1 (Der f 1) allergens to further produce recombinant allergens for future clinical applications in order to eliminate side reactions from crude extracts of mites. Based on GenBank data, we designed primers and amplified the cDNA fragment coding for Der f 1 by nested-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Sequence analysis showed the presence of an open reading frame containing 966 bp that encodes a protein of 321 amino acids. Interestingly, homology analysis showed that the Der p 1 shared more than 87 percent identity in amino acid sequence with Eur m 1 but only 80 percent with Der f 1. Furthermore, phylogenetic analyses suggested that D. pteronyssinus was evolutionarily closer to Euroglyphus maynei than to D. farinae, even though D. pteronyssinus and D. farinae belong to the same Dermatophagoides genus. A total of three cysteine peptidase active sites were found in the predicted amino acid sequence, including 127-138 (QGGCGSCWAFSG), 267-277 (NYHAVNIVGYG) and 284-303 (YWIVRNSWDTTWGDSGYGYF). Moreover, secondary structure analysis revealed that Der f 1 contained an a helix (33.96 percent), an extended strand (17.13 percent), a ß turn (5.61 percent), and a random coil (43.30 percent). A simple three-dimensional model of this protein was constructed using a Swiss-model server. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Alignment and phylogenetic analysis suggests that D. pteronyssinus is evolutionarily more similar to E. maynei than to D. farinae.

Arsenic trioxide reduces the invasive and metastatic properties of nasopharyngeal carcinoma cells in vitro

Nasopharyngeal carcinoma (NPC) is notorious for the metastases, which are in close association with Epstein-Barr virus-encoded latent membrane protein 1 (LMP1). Arsenic trioxide (As2O3) has been shown to induce apoptosis and differentiation in NPC xenografts. Then, can it repress the cancer cells' metastasis potential? To elucidate this issue, the present study was performed. LMP1-negative cell line HNE1 and LMP1-positive cell line HNE1-LMP1 were used as in vitro model. Cells (1 x 10(5)/mL) were cultured with or without 3 æM As2O3 for 48 h. Then the survival cells were collected to investigate their potential of colony formation, attachment, invasion, and migration. Both confocal immunofluorescence staining and Western blot were used to detect the changes of LMP1 expression. The changes of MMP-9 were examined by RT-PCR assay and Western blot. The results were as follow: i) the colony formation inhibition rate (75.41 ± 3.9 percent in HNE1-LMP1 cells vs 37.89 ± 4.9 percent in HNE1 cells), the rate of attachment (HNE1-LMP1 vs HNE1: 56.40 ± 3.5 vs 65.87 ± 5.9 percent), the invasion inhibitory rate (HNE1-LMP1 vs HNE1: 56.50 ± 3.7 and 27.91 ± 2.1 percent), and the migration inhibitory rate (HNE1-LMP1 vs HNE1: 48.70 ± 3.9 vs 29.19 ± 6.27 percent) were all significantly different between the two cell lines (P < 0.01). ii) LMP1 was down-regulated in As2O3-treated HNE1-LMP1 cells. iii) The reduction of MMP-9 was found in As2O3-treated groups, more evident in HNE1-LMP1 cells. Thus, we conclude that As2O3 can reduce metastasis potential of NPC cells, involving inhibition of MMP-9 expression. LMP1 were also reduced in this process and seemed to enhance anti-metastasis activity of As2O3.

A double-blind, placebo-controlled, and randomized study of loratadine (Clarityne) syrup for the treatment of allergic rhinitis in children aged 3 to 12 years.

Allergic rhinitis is a common disease in children, and antihistamines are the key medication. However, traditional tablets are not convenient and lead to low compliance in young children. The aim of this double-blind, placebo-controlled, parallel, randomized study was to evaluate the effectiveness and safety of loratadine syrup for the treatment of children aged 3 to 12 years with allergic rhinitis. Sixty children with allergic rhinitis due to dust mites were enrolled. They were randomized into 2 parallel groups: one group received loratadine syrup 5 mg or 10 mg daily for 3 weeks, and the other group received placebo. The patients returned to special clinics for symptoms evaluation at day 7 and day 21, and the parents were requested to record disease severity daily. Both evaluations, physician's and parents', were recorded with a 4-point scale for 5 symptoms: sneezing, rhinorrhea, nasal congestion, nasal itching and ocular symptoms. Forty-six patients completed the study, 22 in the loratadine group and 24 in the placebo group. At the initial visit, the total symptom score (TSS) in both groups was not significantly different (p = 0.39). The TSS of the loratadine syrup group at day 7 and day 21 was lower than those of the placebo group (p = 0.003, p = 0.06). The daily card scores in the experimental group were also significantly lower than those of the placebo group (week 1, p = 0.014; week 2, p = 0.029; week 3, p = 0.014). No adverse reactions were recorded in both groups. This study revealed that loratadine syrup 5 mg or 10 mg once a day improved symptom scores of children with allergic rhinitis effectively and safely.

Correlation of total and specific serum immunoglobulin E levels with the severity of chronic sinusitis in children.

Chronic sinusitis is frequently associated with allergy and asthma. Previous studies have shown that serum immunoglobulin E (IgE) levels correlate with allergy and asthma in adults. Because the role of allergic inflammation in the severity of chronic sinusitis remains controversial in children, we set out to determine whether a correlation exists between serum levels of total and specific IgE and the severity of chronic sinusitis in children. Forty-four children with chronic sinusitis were enrolled in the study. Computed tomographic scans were reviewed and scored for the severity of sinusitis. All children were mite-sensitive. Serum samples were assayed for total IgE and specific IgE antibodies to mite allergen using a fluoroimmunoassay. Fourteen subjects had extensive sinus disease. There was no significant difference in the average of total and specific IgE between the subjects with extensive and limited disease (p = 0.562 and 0.755, respectively). Thirty-four subjects were diagnosed with asthma. The subjects with extensive sinus disease had a higher prevalence of moderate to severe asthma than the subjects with limited disease (p = 0.006), but there was no significant difference in the average of total and specific IgE between the subjects with different severities of asthma. (p = 0.833 and 0.425, respectively). The data suggests that levels of total or specific IgE do not correlate with severity of chronic sinusitis in children. Nonetheless, the severity of chronic sinusitis and asthma correlate well with each other irrespective of total and specific IgE.

Antibodies in serum of patients with clonorchiasis before and after treatment.

Sera of 31 patients infected with Clonorchis sinensis were examined using fraction 1 antigen by ELISA during a one-year observation. The results of ELISA with Igs, IgG and IgA demonstrated high sensitivity (100%, 100% and 90%) and specificity (100%, 100% and 87%). Sera specific Igs and IgG were significantly decreased in the 3rd month after treatment with praziquantel (25mg/kg body weight in one dose), and IgA was significantly decreased in the 1st month (paired t-test, p < 0.05). No eggs were found in the stool after treatment. Detection of sera specific Igs, IgG and IgA by ELISA was combined with stool examination to evaluate the effect of praziquantel and the completeness of the cure.

Selective IgG subclass deficiencies in patients with recurrent sinopulmonary infections: report of two cases.

Two patients with recurrent sinopulmonary infections and normal total serum immunoglobulin levels were found to have selective deficiencies in IgG subclasses. The serum of one patient contained abnormally low IgG2 and IgG4; and the other was deficient in IgG4. Both patients responded to the treatment with high dose intravenous immunoglobulin. The experiences on these two cases strongly suggest that IgG subclasses should be checked in patients with recurrent sinopulmonary infections in face of normal total immunoglobulins.

Chronic granulomatous disease in two Chinese families.

Two Chinese families with X-linked chronic granulomatous disease (CGD) are reported. The first case was an 11-month-old male baby and the second a 2-month-old male baby. Both patients presented with persistent infections caused by Staphylococcus and Candida since birth. Neutrophil functions were studied in patients and a number of family members. Chemotaxis and phagocytosis were normal in every subject. Slide and spectrophotometric nitroblue tetrazolium (NBT) tests of both patients were abnormal and remained unchanged in spite of treatment with ascorbic acid, levamisole, sulfamethoxazole, trimethoprim and isoniazide. Mothers were proved to be carriers as evidenced by the presence of both normal and CGD phagocytes in the slide NBT test. During the 2-month follow-up period, the percentage of normal phagocytes from the mother of case 1 varied from 12% to 73%, which correlated with the fluctuation of spectrophotometric NBT value. The slide NBT test of the mother of case 2 was nearly normal in face of the presence of CGD phagocytes. Both carrier mothers were healthy and asymptomatic.