Cloning and gene function of dicarboxylate-tricarboxylate carrier protein in Gastrodia elata / 中国中药杂志

The gene GeDTC encoding the dicarboxylate-tricarboxylate carrier protein in Gastrodia elata was cloned by specific primers which were designed based on the transcriptome data of G. elata. Bioinformatics analysis on GeDTC gene was carried out by using ExPASY, ClustalW, MEGA, etc. Positive transgenic plants and potato minituber were obtained by virtue of the potato genetic transformation system. Agronomic characters, such as size, weight, organic acid content, and starch content, of potato minituber were tested and analyzed and GeDTC gene function was preliminarily investigated. The results showed that the open reading frame of GeDTC gene was 981 bp in length and 326 amino acid residues were encoded, with a relative molecular weight of 35.01 kDa. It was predicted that the theoretical isoelectric point of GeDTC protein was 9.83, the instability coefficient was 27.88, and the average index of hydrophilicity was 0.104, which was indicative of a stable hydrophilic protein. GeDTC protein had a transmembrane structure and no signal peptide and was located in the inner membrane of mitochondria. The phylogenetic tree showed that GeDTC was highly homologous with DTC proteins of other plant species, among which GeDTC had the highest homology with DcDTC(XP＿020675804.1) in Dendrobium candidum, reaching 85.89%. GeDTC overexpression vector pCambia1300-35Spro-GeDTC was constructed by double digests, and transgenic potato plants were obtained by Agrobacterium-mediated gene transformation. Compared with the wild-type plants, transgenic potato minituber harvested by transplanting had smaller size, lighter weight, lower organic acid content, and no significant difference in starch content. It is preliminarily induced that GeDTC is the efflux channel of tricarboxylate and related to the tuber development, which lays a foundation for further elucidating the molecular mechanism of G. elata tuber development.

Advances on the production of organic acids by yeast / 生物工程学报

Organic acids are organic compounds that can be synthesized using biological systems. They often contain one or more low molecular weight acidic groups, such as carboxyl group and sulphonic group. Organic acids are widely used in food, agriculture, medicine, bio-based materials industry and other fields. Yeast has unique advantages of biosafety, strong stress resistance, wide substrate spectrum, convenient genetic transformation, and mature large-scale culture technology. Therefore, it is appealing to produce organic acids by yeast. However, challenges such as low concentration, many by-products and low fermentation efficiency still exist. With the development of yeast metabolic engineering and synthetic biology technology, rapid progress has been made in this field recently. Here we summarize the progress of biosynthesis of 11 organic acids by yeast. These organic acids include bulk carboxylic acids and high-value organic acids that can be produced naturally or heterologously. Finally, future prospects in this field were proposed.

Sodium butyrate in post-hatch feed for broiler chicks / [Uso de butirato de sódio em rações pós-eclosão para pintos de corte]

This study was designed to determine adequate levels for sodium butyrate inclusion in pre-hatching and pre-starter feed in order to minimize the negative effects of post-hatch delayed placement on broiler chicks. Newly-hatched chicks were allotted in a completely randomized design, with five treatments, each comprising five replicates of ten birds each. Five dietetic levels of sodium butyrate (control, 0.050, 0.075, 0.100 and 0.200%) were used in the pre-starter feed offered to the chicks in the transporting box and during the pre-starter phase. Performance, yolk sac retraction, plasma glucose concentration, weight and histomorphometry of the small intestine were evaluated after 24 hours of feed access and at 7 days of age. A metabolic trial was performed when the chicks were seven to ten days of age. Supplementation of 0.1% sodium butyrate increased the development of broiler chicks' intestinal villi at seven days of age but was not able to improve performance in the pre-starter phase. Supplementation with sodium butyrate in a diet offered in the transportation box does not improve broiler performance in the pre-starter phase. Supplementation of up to 0.16% is recommended, in order to improve the metabolizability of the ether extract for broilers at seven days of age.(AU)

Objetivou-se, com este estudo, encontrar níveis adequados de inclusão de butirato de sódio em ração pós-eclosão e pré-inicial, buscando minimizar os efeitos negativos do jejum de pintos de corte. Os animais foram distribuídos em delineamento inteiramente ao acaso, com cinco tratamentos e cinco repetições de 10 aves cada. Foram utilizadas cinco suplementações de butirato de sódio (controle; 0,05; 0,075; 0,100 e 0,200%) na ração pré-inicial, fornecidas aos pintos na caixa de transporte e durante a fase pré-inicial. Após 24 horas de acesso ao alimento e aos sete dias de idade, foram avaliados desempenho, retração do saco vitelino, concentração de glicose plasmática, peso e histomorfometria do intestino delgado. De sete a 10 dias de idade, realizou-se ensaio de metabolizabilidade. A suplementação de 0,10% de butirato de sódio aumentou o desenvolvimento das vilosidades intestinais dos pintos de corte aos sete dias de idade, mas não foi capaz de melhorar o desempenho na fase pré-inicial. A suplementação com butirato de sódio em dieta oferecida na caixa de transporte não melhora o desempenho dos frangos na fase pré-inicial. Recomenda-se suplementação de até 0,16%, a fim de melhorar a metabolizabilidade do extrato etéreo para frangos de corte aos sete dias de idade.(AU)

Determination of Salt Forms of New Psychoactive Substances by Ion Chromatography / 法医学杂志

Objective To establish an ion chromatography method for the salt form determination of new psychoactive substances （NPS）. Methods The method of conducting qualitative and quantitative analysis of six types of organic acid ions （acetate ion, tartrate ion, maleate ion, oxalate ion, fumarate ion, citrate ion） and five types of inorganic anions （fluoride ion, chloride ion, nitrate ion, sulfate ion, phosphate ion） in NPS sample by ion chromatography was developed. The salt forms of 222 seized NPS samples （103 samples with synthetic cannabinoids, 81 samples with cathinones, 44 samples with phenylethylamines, 12 samples with tryptamines, 7 samples with phencyclidines, 6 samples with piperazines, 2 samples with aminoindenes, 26 samples with fentanyls and 43 samples with other types of NPS） were analyzed by this method. Results Each anion had good linearity in the corresponding linear range, the correlation coefficients （r） were greater than 0.999, the limits of detection were 0.01-0.05 mg/L, and the limits of quantitative were 0.1-0.5 mg/L. Except that 5F-BEPIRAPIM was hydrochloride, the salt forms of the other 102 synthetic cannabinoids were all base. The salt form of 81 cathinone samples, 44 phenylethylamine samples, 7 phencyclidine samples and 2 aminoindene samples were all hydrochloride. The salt forms of tryptamine samples included base, hydrochloride, fumarate and oxalate. The salt forms of piperazine samples included base and hydrochloride. The salt forms of fentanyl samples and samples of other types included base, hydrochloride and citrate. Conclusion Ion chromatography is a simple, accurate and efficient method for determining the salt form of NPS samples, which makes the qualitative and quantitative conclusions of NPS more scientific and rigorous.

Advances in the metabolic engineering for the production of tetracarbon organic acids / 生物工程学报

Tetracarbon organic acids are important platform chemicals that are widely used in the food, chemical, medicine, material industries and agriculture. Compared with the traditional petrochemical process, the production of tetracarbon organic acids by microbial fermentation is more promising due to milder reaction conditions, greener process and better environmental compatibility. This review summarizes the biosynthetic pathways and metabolic mechanisms for the production of tetracarbon organic acids, and illustrates recent advances, challenges, and future perspectives in the production of tetracarbon organic acids by naturally selected or purposefully engineered strains.

Silage from heart-of-palm waste produced from Alexander palm tree / Ensilaje de residuos de palmito producidos a partir de Palma Real / Silagem de resíduos de palmito produzidos a partir da Palmeira Real

Abstract Background: Production of heart-of-palm from Alexander palm tree generates a waste that could be used as silage for feeding ruminants. Objective: To evaluate the chemical composition, fermentation parameters, and quality of silage made from waste of heart-of-palm production using Alexander palm. Methods: Three types of silage made from waste of heart-of-palm production were evaluated: 1) leaf silage, 2) sheath silage, and 3) compound (leaf + sheath) silage. The processed waste was packed in experimental silos and distributed in a completely randomized design with 10 replications. Means were compared pairwise using Tukey's test at the 5% significance level. Results: Leaf silage had the highest pH (p<0.05) and the lowest ammonia nitrogen (p<0.05) in relation to the other silages. No difference was observed (p>0.05) for lactic and butyric acids, whereas acetic acid was higher (p<0.05) in the sheath and compound silages. Dry matter and crude protein contents were the highest (p<0.05) in leaf silage. Neutral detergent fiber was higher (p<0.05) in sheath silage, followed by leaf and compound silages, respectively. The highest lignin content (p<0.05) was observed in sheath silage. Conclusions: Based on the fermentative parameters, waste from heart-of-palm from Alexander palm tree has potential for silage production. However, differences in silage composition at the time of feeding ruminants should be considered.

Resumen Antecedentes: Los residuos de la producción de palmito de Palma Real podrian utilizarse como ensilaje para alimentación de rumiantes. Objetivo: Determinar la composición, parámetros químicos de fermentación, y calidad del ensilaje elaborado a partir de residuos procedentes de la producción de palmito de Palma Real. Métodos: Tres tipos de ensilaje hechos con los residuos generados a partir de la producción de palmito fueron evaluados: 1) ensilaje de hojas, 2) ensilaje de vainas, y 3) ensilaje compuesto (hojas + vainas). Los residuos tratados fueron empacados en silos experimentales y distribuidos en un diseño completamente al azar con 10 repeticiones. Las medias fueron comparadas mediante la prueba de Tukey, con un nivel de significancia del 5%. Resultados: El ensilaje de hojas mostró el pH más alto (p<0,05) y el nivel más bajo de nitrógeno amoniacal (p<0,05) en comparación con los otros ensilajes. No se observaron diferencias (p>0,05) en los ácidos láctico y butírico, mientras que el ácido acético fue mayor en el ensilaje de vainas y en el compuesto en relación con el otro ensilaje. La materia seca (MS) y el contenido de proteína cruda (PC) fueron mayores (p<0,05) en el ensilaje de hojas. Los contenidos de fibra detergente neutra fueron mayores (p<0,05) en los ensilajes de vainas, seguido por los ensilajes de hojas y compuesto, respectivamente. El contenido de lignina fue más alto (p<0,05) en el ensilaje de vainas. Conclusiones: Según los parámetros fermentativos, los residuos de la producción de palmito de Palma Real tienen potencial para la producción de ensilaje. Sin embargo, se deben considerar las diferencias en la composición de los ensilajes al momento de alimentar rumiantes.

Resumo Antecedentes: Resíduos da produção de palmito da palmeira Real são um problema para a agroindústria devido sua falta de utilização e o seu uso na forma de silagem pode ser uma alternativa na alimentação de ruminantes. Objetivo: Avaliar a composição química e os parâmetros fermentativos, e verificar a qualidade da silagem produzida a partir do resíduo da produção de palmito da Palmeira Real Australiana. Métodos: Foram avaliadas três tipos de silagem de resíduo da produção de palmito da Palmeira Real: silagem de folha, silagem de bainha e silagem da composta (folha + bainha). O resíduo processado foi acondicionado em silos experimentais e distribuídos em um delineamento inteiramente casualizado com 10 repetições e as médias comparadas pelo teste de Tukey a 5% de significância. Resultados: A silagem de folha apresentou maior pH (p<0,05) em relação as demais silagens, enquanto para o nitrogênio amoniacal (p<0,05), apresentou menor teor em relação as demais silagens. Não observou-se diferença (p>0,05) para os ácidos lático e butírico, enquanto o ácido acético foi maior para as silagens de bainha e composta em relação as demais silagens. O teor de matéria seca (MS), proteína bruta (PB) foi maior (p<0,05) na silagem de folha em relação as demais silagens. A fibra em detergente neutro analisada estimada apresentou maiores valores (p<0,05) para a silagem de bainha, seguida pela silagem de folhas e silagem composta, respectivamente. O teor de lignina foi maior (p<0,05) para a silagem de bainha. Conclusões: De acordo com os parâmetros fermentativos, os resíduos da produção de palmito da Palmeira Real apresentam potencial para a produção de silagem. No entanto, as diferenças na composição das silagens devem ser consideradas quando oferecidas para animais ruminantes.

Mechanisms and regulation of aluminum-induced secretion of organic acid anions from plant roots / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Aluminum (Al) is the most abundant metal element in the earth's crust. On acid soils, at pH 5.5 or lower, part of insoluble Al-containing minerals become solubilized into soil solution, with resultant highly toxic effects on plant growth and development. Nevertheless, some plants have developed Al-tolerance mechanisms that enable them to counteract this Al toxicity. One such well-documented mechanism is the Al-induced secretion of organic acid anions, including citrate, malate, and oxalate, from plant roots. Once secreted, these anions chelate external Al ions, thus protecting the secreting plant from Al toxicity. Genes encoding the citrate and malate transporters responsible for secretion have been identified and characterized, and accumulating evidence indicates that regulation of the expression of these transporter genes is critical for plant Al tolerance. In this review, we outline the recent history of research into plant Al-tolerance mechanisms, with special emphasis on the physiology of Al-induced secretion of organic acid anions from plant roots. In particular, we summarize the identification of genes encoding organic acid transporters and review current understanding of genes regulating organic acid secretion. We also discuss the possible signaling pathways regulating the expression of organic acid transporter genes.

Cloning and expression of key enzyme genes for chlorogenic acid synthesis in Chrysanthemum morifolium / 中草药

To investigate the effects of the expression of coumaroylquinate 3’-monooxygenase (C3’H) and shikimate O-hydroxycinnamoyltransferase (HCT) in the chlorogenic acid-producing pathway and active ingredients in Chrysanthemum morifolium under flooding stress, we cloned two C3’H genes which were CmC3’H1 and CmC3’H2 and two HCT genes which were CmHCT1 and CmHCT2 by the RT-PCR from Hangju and conducted bioinformatics analysis. During the flower bud differentiation stage, we flooded the C. morifolium and then used β-actin as the reference gene to detect the relative expression of the four genes by the qRT-PCR. Finally, the content of downstream products and other indicators of these four genes in C. morifolium were measured by HPLC. We obtained the four genes’ complete open reading frame and predicted the relative molecular mass of the amino acid sequence and the theoretical isoelectric point (pI). And the protein tertiary structure models were constructed. The qRT-PCR results showed that flooding the C. morifolium for 3 days during the flower bud differentiation stage resulted in significant expression changes of the four genes at different growth stages. The results of HPLC showed that chlorogenic acid, the downstream product catalyzed by the C3’H and the HCT, was significantly higher than that in the control group. It was also found that the content of luteoloside and 3,5-O-di-caffeoylquinic acid was also significantly higher than those of the control group. Therefore, C. morifolium regulates the synthesis of downstream products by regulating the expression of the four genes under flooding stress, thereby responding to flooding stress. And the flooding stress during flower bud differentiation can significantly enhance the accumulation of active ingredients of C. morifolium.

Mechanisms and regulation of aluminum-induced secretion of organic acid anions from plant roots / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Aluminum (Al) is the most abundant metal element in the earth’s crust. On acid soils, at pH 5.5 or lower, part of insoluble Al-containing minerals become solubilized into soil solution, with resultant highly toxic effects on plant growth and development. Nevertheless, some plants have developed Al-tolerance mechanisms that enable them to counteract this Al toxicity. One such well-documented mechanism is the Al-induced secretion of organic acid anions, including citrate, malate, and oxalate, from plant roots. Once secreted, these anions chelate external Al ions, thus protecting the secreting plant from Al toxicity. Genes encoding the citrate and malate transporters responsible for secretion have been identified and characterized, and accumulating evidence indicates that regulation of the expression of these transporter genes is critical for plant Al tolerance. In this review, we outline the recent history of research into plant Al-tolerance mechanisms, with special emphasis on the physiology of Al-induced secretion of organic acid anions from plant roots. In particular, we summarize the identification of genes encoding organic acid transporters and review current understanding of genes regulating organic acid secretion. We also discuss the possible signaling pathways regulating the expression of organic acid transporter genes.

A report on genetic analysis for 3 cases of glutaric academia type Ⅰ / 临床检验杂志

Objective@#To verify the diagnosis of highly suspected glutaric academia type I for the cases found in neonatal screening and conduct related genetic analysis. Methods: In this research the clinical data of the children with glutaric academia type I were collected, and the diagnostic panels of inherited metabolism diseases with gene capture high-throughput sequencing technology were applied to perform genetic diagnosis in suspected cases. Sanger sequencing technology was also used to verify the genes of the members in this family. In addition, we searched a large number of relevant literatures for genetic analysis. @*Methods@#In this research the clinical data of the children with glutaric academia type I were collected, and the diagnostic panels of inherited metabolism diseases with gene capture high-throughput sequencing technology were applied to perform genetic diagnosis in suspected cases. Sanger sequencing technology was also used to verify the genes of the members in this family. In addition, we searched a large number of relevant literatures for genetic analysis. @*Results@#All the 3 cases were found to have complex heterozygous mutation sites of GCDH gene by gene sequencing technology. The mutation sites were c.109_110delCA and c.416C＞G in the first case, c.892G＞A and c.261_506-433delinsATA in the second case and c.1235C＞A and c.1244-2A＞C in the last case. Among them, c.261_506-433delinsATA and c.109_110delCA should be completely newly identified and never reported in literatures. All the mutation sites were verified to be inherited from their parents. @*Conclusion@#Next-generation sequencing technology can contribute to confirming the diagnosis of glutaric academia type I and provide reliable evidence for appropriate treatment and genetic counseling of this disease.

Use of succinic & oxalic acid in reducing the dosage of colistin against New Delhi metallo-?-lactamase-1 bacteria

Background & objectives: New Delhi metallo-?-lactamase 1 (NDM-1) cleaves the beta-lactam ring, and confers bacterial resistance against most of the beta-lactam antibiotics, except tigecycline and colistin. Among these two antibiotics, colistin is considered toxic, and therefore, its clinical use and dosage need cautious approach. In the present study, six organic acids were screened individually and in combination of two acids for their effectiveness against NDM-1 Escherichia coli and a combination of colistin and oxalic or succinic acid was tested to find out the potential of combination therapy for reducing the dose of toxic colistin. Methods: Antibacterial activity of the organic acid and their combinations was tested by disc diffusion method against NDM-1 E. coli, and minimum inhibitory concentration (MIC) was determined by broth dilution method. Synergistic effect between organic acids and colistin was tested by checkerboard method. Results: Oxalic acid showed the highest zone of inhibition (15�mm) followed by succinic acid, tartaric acid, fumaric acid, citric acid and malic acid. The combination of two acids did not increase the zone of inhibition significantly. MIC was found to be the lowest with oxalic acid and succinic acid (320 ?g/ml). In the presence of 160 ?g/ml oxalic acid or succinic acid, MIC of colistin was reduced from 8 to 4 ?g/ml, indicating synergistic effect. Interpretation & conclusions: Our findings showed that combination therapy using colistin and oxalic acid or succinic acid might find safe clinical application of this antibiotic in controlling infections due to NDM-1 bacteria.

Report on the External Quality Assessment Scheme for Metabolite Testing in Korea (2016–2017)

External quality assessment (EQA) trials of conventional newborn screening tests for phenylketonuria, galactosemia, congenital adrenal hyperplasia, maple syrup urine disease, homocystinuria, and congenital hypothyroidism, as well as extended newborn screening tests using tandem mass spectrometry, were performed twice in 2016 and 2017. A total of 44 specimens in the form of dried blood spots were distributed in each trial to 16 laboratories. The response rate of these laboratories was 100%. The mean, standard deviation, coefficient of variation, median, and cut-offs were evaluated for each analyte in the newborn screening tests. EQA trials for the analyses of methylmalonic acid, vanillylmandelic acid, catecholamines, metanephrines, organic acids, and amino acids were also performed. A well-designed EQA program and continuous education would improve the performance of metabolite testing.

Mimicking Ketonuria in the Ketogenesis Defect 3-Hydroxy-3-MethylglutarylCoenzyme A Lyase Deficiency: An Artefact in the Analysis of Urinary Organic Acids

Abstract 3-Hydroxy-3-methylglutaryl-coenzyme A lyase (HMGCL, HMGCL) deficiency is a rare inborn error of ketogenesis. Even if the ketogenic enzyme is fully disrupted, an elevated signal for the ketone body acetoacetic acid is a frequent observation in the analysis of urinary organic acids, at least if derivatization is performed by methylation. We provide an explanation for this phenomenon and trace it back to degradation of the derivatized 3-hydroxy-3-methylglutaric acid and high temperature of the injector of the gas chromatograph.

Rapid identification of constituents from different Ginkgo biloba preparations by high resolution mass spectrometry and metabolomics technology / 中国药科大学学报

@#A qualitative analysis was developed to identify different ingredients from three Ginkgo biloba preparations(including Yinxingye Diwan, extract of Ginkgo biloba leaves tablets and Yinxingtongzhi Diwan)based on high resolution mass spectrometry and metabolomics technology. An XSELECT HSS T3(4. 6 mm×150 mm, 3. 5 μm)column was used for separation, with the mobile phases consisting of acetonitrile and water containing 0. 1% formic acid. The column temperature was set at 40 °C. Negative ion mode was used for mass spectrometric data acquisition. Through partial least squares projection to latent structure-discriminant analysis(PLS-DA), we firstly found the different compounds among different Ginkgo biloba preparations. Subsequently, the compounds displaying different abundance levels via database searching and literature matching were identified. Finally, we identified 21 different compounds between the extract of Ginkgo biloba leaves tablets and Yinxingye Diwan, which mostly belong to the organic acids and flavonols families. Quantitative analysis showed that the ingredients which had higher abundance in extract of Ginkgo biloba leaves tablets were mainly organic acids, whereas those exhibiting higher levels in Yinxingye Diwan are flavonols. We also identified 12 different ingredients between the Yinxingye Diwan and Yinxingtongzhi Diwan, which were flavonols, sciadopitysin and organic acids. The results of this study are useful for studying different chemical constituents from distinct Ginkgo biloba preparations.

Principal component determination and analysis of urine organic acid metabolism spectrum of two diseases / 基础医学与临床

Objective Analysis of 22 kinds of organic acid metabolites in urine samples of patients with myasthenia gravis, mitochondrial myopathy and of healthy controls was performed so to provide data and basis for clinical screening .Methods The principal component and the corresponding principal component equation were obtained , the physical and chemical significance of the principal component was explained .Results The cumulative contri-bution rate of the first five principal components reached 86.89%, was identified as the main component , then es-tablished the principal component function expression , and analyzed the relationship between the principal compo-nent and the original variable .It was found that the phenyl saturated acid might be a potential biomarker of the two diseases , and the hippuric acid was an early warning bio-marker of the two diseases .Conclusions Urine organic acid metabolic profile principal component analysis is helpful to find biomarker of disease and may support clinical diagnosis basis .

Clinical analysis of methylmalonic acidemia in 21 children / 临床儿科杂志

Objective To explore the clinical features of methylmalonic acidemia (MMA) in children admitted to the pediatric intensive care unit, to help improve our understanding of MMA. Methods The clinical data of 21 patients with MMA admitted to our PICU from December 2012 to August 2016 were analyzed. Diagnosis were confirmed by gas chromatography-mass spectrometry, GC/MS. Results twenty-four of 158 suspected cases were confirmed as having organic acidemia diseases including 21 cases of MMA, one case of propionic acidemia, one case of urea cycle disorders, and one case of glutaric acidemia. The main clinical manifestations were feeding difficulty, malnutrition (13 cases), developmental retardation (12 cases), lethargy (10 cases), tricuspid severe reflux and pulmonary hypertension (1 case), hydrocephaly (5 cases), muscular dystonia (three cases with hypertonia, and four with hypotonia), convulsion (7 cases), apnea, sobbing respiration (10 cases), chromatosis (6 cases), anemia (13 cases), edema (6 cases), thrombocytopenia (6 cases), hematuria and proteinuria (2 cases). Five cases gave up therapy before diagnosis was made. Sixteen cases received the treatment with Vitamin B12 and supplementation of L-carnitine. Seven cases gave up after treatment without effect or deterioration of condition. Eight cases were vitamin B12-responsive, and one case was vitamin B12-nonresponsive. The follow-up for a period ranging from three months to two years, among eight vitamin B12-responsive cases, 6 cases showed a favorable outcome with apparent improvement, one case had no symptom and one patient died from severe pneumonia. Vitamin B12-nonresponsive case was still alive. Conclusions The clinical manifestations of MMA are non-specific. Urine organic acid analysis is critical to early diagnosis of MMA in high-risk patients. Timely diagnosis and appropriate long-term treatment are essential to improve the prognosis of the disease.

Screening of Urinary Organic Acid in Children with High Risk of Inherited Metabolic Diseases / 中国康复理论与实践

Objective To explore the application of gas chromatography mass spectrometry (GC/MS) in children with high risk of inher-ited metabolic disease. Methods From March, 2010 to November, 2015, 119 children suspected with inherited metabolic disease were in-cluded. The urinary organic acid was detected with GC/MS, and related diseases were screened. Results Seventeen children (14.29%) were positive with inherited metabolic disease, in which 16 cases (94.12%) manifested with development retardation. 20 children (16.81%) were probable positive. Conclusion GC/MS is effective in screening children with high risk of inherited metabolic disease, which can provide ba-sis for further diagnosis.

Effects of Total Organic Acid of Thladiantha dubia Fruit on Coagulation Time and Hemorheology of Rats with Cold Coagulation and Blood Stasis / 中国药房

OBJECTIVE:To observe the effects of total organic acid of Thladiantha dubia fruit(TOATF)on coagulation time and hemorheology of rats with cold coagulation and blood stasis. METHODS:60 rats were randomly divided in normal control group (water),model control group (water),aspirin group (positive drug,50 mg/kg) and TOATF low-dose,medium-dose and high-dose groups(50,100,200 mg/kg)with 10 rats in each group. Except for normal control group,cold coagulation and blood stasis model was induced by 4 ℃ water bath and subcutaneous injection of adrenaline hydrochloride,and then given correspon-dence medicine intragastrically,once a day,for consecutive 14 d. 24 h after last administration,blood samples were collected from aorta abdominalis. The coagulating time(CT),erythrocrit(HCT),plasma viscosity(PV),prothrombin time(PT),thrombin time (TT),activated partial thromboplastin time (APTT),platelet aggregation rate (PAR) and whole blood low-shear,middle-shear and high-shear viscosity were measured and blood sedimentation equation K value was calculated. RESULTS:Compared with nor-mal control group,CT,PT,TT and APTT of model control group were shortened,and HCT,PV,blood sedimentation equation K value,PAR and whole blood low-shear,middle-shear and high-shear viscosity increased(P<0.05 or P<0.01). Compared with model control group,CT,PT and APTT prolonged in aspirin group and TOATF medium-dose and high-dose groups,and PAR de-creased;PT of treatment groups prolonged,while HCT,PV,blood sedimentation equation K value and whole blood low-shear and middle-shear viscosity decreased(P<0.05 or P<0.01). CONCLUSIONS:TOATF has obvious improvement effects on anticoagula-tion and hemorheology in rats with cold coagulation and blood stasis.

Simultaneous determination of 14 organic acids in Shenfu injection by hydrophilic interaction chromatography-tandem mass spectrometry / 中国中药杂志

Organic acids are widely distributed in plants and related products, and participate in a wide range of metabolic pathways (e.g. tricarboxylic acid cycle), showing diverse pharmacological activities. As a widely used Chinese patent medicine, its adverse reactions are often reported. Therefore, we should further clarify the chemical components of Shenfu injection, and prepare strict quality standards to ensure the safety and effectiveness of its clinical use. Shenfu injection is prepared from red ginseng (steamed roots of Panax ginseng) and black prepared lateral roots of Aconitum carmichaelii (Heishunpian) by using modern extraction process, and organic acids are regarded as one of its main components. In current study, a hydrophilic interaction chromatography (HILIC) coupled with mass spectrometric method (HILIC-LC-MS) was developed and validated for the simultaneous determination of 14 organic acids, including cinnamic acid, ferulic acid, 4-hydroxylbenzoic acid, L-(+)-lactic acid, adipic acid, fumaric acid, caffeic acid, succinic acid, maleic acid, malonic acid, D-malic acid, (-)-shikimic acid, D-tartaric acid, and quinic acid in Shenfu injection. Satisfactory retention and separation were achieved for all organic acids on HILIC chromatographic column. Except cinnamic acid (231 μg•L⁻¹), lactic acid (113 μg•L⁻¹) and malonic acid (32.5 μg•L⁻¹), the limit of quantitation for the remaining 11 compounds were less than 10 μg•L⁻¹. D-Malic acid, malonic acid, quinic acid, L-(+)-lactic acid, and cinnamic acid were observed to have higher contents in Shenfu injection (>1.89 mg•L⁻¹), whereas caffeic acid and adipic acid were undetectable in all batches. Above all, the developed method is suitable for the simultaneous determination of organic acids in Shenfu and some other traditional Chinese medicine injections.

Simultaneous determination of multiple constituents in scrophulariae radix from different habitats and commercial herbs by UPLC-QTRAP-MS/MS / 中国药学杂志

OBJECTIVE: To develop a method for the determination of iridoid, phenylpropanoid glycosides, and organic acids in Scrophulariae Radix from different habitats and commercial herbs by UPLC-QTRAP-MS/MS. METHODS: The analysis was carried out on a BDS HYPERSIL C18 column (4.6 mm×250 mm, 5 μm) with elution by mobile phase of acetonitrile-water at a flow rate of 1.0 mL·min-1. The column temperature was maintained at 35℃. The target compounds were analyzed by the negative ion multiple reaction monitoring (MRM) mode. RESULTS: Twelve multiple constituents showed good linearity (r>0.9994) in the range of the tested concentration. The average recoveries of the 12 components were 99.59%-101.24% with relative standard deviations of 0.93%-1.60%. CONCLUSION: The established method is accurate and precise, which provides a reliable and effective technique for the quality evaluation of Scrophulariae Radix.