Urinary metabolic insights into host-gut microbial interactions in healthy and IBD children.

AIM: To identify metabolic signatures in urine samples from healthy and inflammatory bowel disease (IBD) children. METHODS: We applied liquid chromatography and gas chromatography coupled to targeted mass spectrometry (MS)-based metabolite profiling to identify and quantify bile acids and host-gut microbial metabolites in urine samples collected from 21 pediatric IBD patients monitored three times over one year (baseline, 6 and 12 mo), and 27 age- and gender-matched healthy children. RESULTS: urinary metabolic profiles of IBD children differ significantly from healthy controls. Such metabolic differences encompass central energy metabolism, amino acids, bile acids and gut microbial metabolites. In particular, levels of pyroglutamic acid, glutamic acid, glycine and cysteine, were significantly higher in IBD children in the course of the study. This suggests that glutathione cannot be optimally synthesized and replenished. Whilst alterations of the enterohepatic circulation of bile acids in pediatric IBD patients is known, we show here that non-invasive urinary bile acid profiling can assess those altered hepatic and intestinal barrier dysfunctions. CONCLUSION: The present study shows how non-invasive sampling of urine followed by targeted MS-based metabonomic analysis can elucidate and monitor the metabolic status of children with different GI health/disease status.

[Determination of mycotoxins in pu-erh tea, black tea, and green tea samples].

To establish a robust method for the determination of mycotoxins in tea samples, and to provide means for the quality and safety control of tea products. Samples of 20 tea products acquired from international market were extracted by organic solvents (acetonitrile containing 0.1% formic acid) or hot water, respectively. The extracts were analyzed by UPLC-MS/MS.A good linear regression was achieved in a range of 39.1 to 5 000 ng•L⁻¹ for aflatoxin B1 (AFB1) and aflatoxin G1 (AFG1), 117 to 15 000 ng•L⁻¹ for aflatoxin B2 (AFB2) and aflatoxin G2 (AFG2), 2.44 to 313 ng•L⁻¹ for fumonisin B1 (FB1), fumonisinB2 (FB2) and fumonisin B3 (FB3), and 3 125 to 5 000 ng•L⁻¹ for deoxynivalenol, with recovery rates between 85.7% and 99.6%. The coefficient of the linear equation for all standards was greater than 0.999 0, and the RSD value was less than 10%. Mycotoxins were detected in several tea samples using the two extraction methods but with different outcomes. The levels of mycotoxins detected ranging from 0.15 to 7.31 µg•kg⁻¹ were well below the State or US FDA regulation limits of mycotoxins in food products. Both methods are simple, accurate, and sensitive, and thus, suitable for the quantitative determination of mycotoxins in different food products. The method with the 80 ℃ hot-water extraction is more appropriate to determine the trace amounts of mycotoxins in tea leaves that are likely to be present in brewed tea liquor, while organic solvent method is more suitable for the detection of mycotoxins in ingestible foods.

Molecular Diversity of Sapovirus Infection in Outpatients Living in Nanjing, China (2011-2013).

Aim. To gain insight into the molecular diversity of sapovirus in outpatients with acute gastroenteritis in Nanjing, China. Methods. The specimens from outpatients clinically diagnosed as acute gastroenteritis were detected by real-time PCR; RT-PCR was then performed to amplify part of VP1 sequences. The PCR products were cloned into pGEM-T Easy vector and bidirectionally sequenced. All sequences were edited and analyzed. A phylogenetic tree was drawn with the MEGA 5.0 software. Results. Between 2011 and 2013, 16 sapovirus positive cases were confirmed by real-time PCR. The infected cases increased from two in 2011 and six in 2012 to eight in 2013. The majority was children and the elderly (15, 93.75%) and single infections (15, 93.75%). Of the 16 real-time positive specimens, 14 specimens had PCR products and the analysis data of the 14 nucleic sequences showed that there was one GI genogroup with four genotypes, two GI.2 in 2011, three GI.2, and one GI.1 in 2012 and one GI.2, three GI.1, two GI.3, and two GI.5 in 2013. Conclusion. Our data confirmed continuous existing of GI genogroup and GI.2 genotype from 2011 to 2013 in Nanjing and the successive appearance of different genotypes from outpatients with gastroenteritis.

Epidemiological and clinical characteristics and risk factors for death of patients with avian influenza A H7N9 virus infection from Jiangsu Province, Eastern China.

BACKGROUND: A novel avian influenza A (H7N9) virus has caused great morbidity as well as mortality since its emergence in Eastern China in February 2013. However, the possible risk factors for death are not yet fully known. METHODS AND FINDINGS: Patients with H7N9 virus infection between March 1 and August 14, 2013 in Jiangsu province were enrolled. Data were collected with a standard form. Mean or percentage was used to describe the features, and Fisher's exact test or t-test test was used to compare the differences between fatal and nonfatal cases with H7N9 virus infection. A total of 28 patients with H7N9 virus infection were identified among whom, nine (32.1%) died. The median age of fatal cases was significant higher than nonfatal cases (P<0.05). Patients with older age were more strongly associated with increased odds of death (OR = 30.0; 95% CI, 2.85-315.62). Co-morbidity with chronic lung disease and hypertension were risk factors for mortality (OR = 14.40; 95% CI, 1.30-159.52, OR = 6.67; 95% CI, 1.09-40.43, respectively). Moreover, the presence of either bilateral lung inflammation or pulmonary consolidation on chest imaging on admission was related with fatal outcome (OR = 7.00; 95%CI, 1.10-44.61). Finally, dynamic monitoring showed that lymphopenia was more significant in fatal group than in nonfatal group from day 11 to week five (P<0.05). The decrease in oxygenation indexes were observed in most cases and more significantly in fatal cases after week three (P<0.05), and the value of nearly all fatal cases were below 200 mmHg during our evaluation period. CONCLUSIONS: Among cases with H7N9 virus infection, increased age accompanied by co-morbidities was the risk of death. The severity of lung infection at admission, the persistence of lymphocytopenia, and the extended duration of lower oxygenation index all contributed to worsened outcomes of patients with H7N9 virus infection.

[Analysis of matrine and oxymatrine in Kushensu injection by UPLC/Q-TOF-MS].

OBJECTIVE: To develop a method for the determination of matrine and oxymatrine in Kushensu injection by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS). METHOD: The analysis was performed on a Acquity UPLC BEH C18 column (2.1 mm x 100 mm, 1.7 microm) with the gradient elution of 10 mmol x L(-1) amine acetate (pH 8, adjusted by aqueous ammonia) and methanol. The flow rate was set as 0.30 mL x min(-1). The column temperature was maintained at 30 degrees C. The peaks were detected at 210 nm and the injection volume was 2 microL. RESULT: The calibration curves showed good linearity within the test ranges of 0.084-3.36 microg for matrine and 0.086-3.44 microg for oxymatrine, respectively. The mean recoveries were 103.2%, 98.7%, and the RSD were 1.5%, 1.2%, respectively. CONCLUSION: The established method is simple, rapid and sensitive, can be used for the quality control of matrine and oxymatrine in Kushensu injection through the manufacturing process and clinical implement.

[Quality control of non-alkaloids from Radix Sophorae tonkinensis].

OBJECTIVE: To establish a method for quality control of non-alkaloids from Radix Sophorae tonkinensis. METHODS: Sugar, organic-acid, polyphenolic and saponin were identified by TLC. The contents of Sugar, polyphenolic and saponin were determined by spectrophotometry and the content of organic-acids was determined by acid-based titration. RESULTS: The contents of sugar, polyphenolic and saponin were 54.37%, 4.74%, 4.40%, respectively. The average content of organic-acid was 9.40%. CONCLUSIONS: The established method is simple, accurate, reproducible and suitable for the quality control of non-alkaloids from Radix Sophorae tonkinensis.

Analysis of dencichine in Panax notoginseng by gas chromatography-mass spectrometry with ethyl chloroformate derivatization.

Dencichine (beta-N-oxalyl-l-alpha,beta-diaminopropionic acid) is a haemostatic agent present in well-known traditional Chinese medicinal herbs such as Panax notoginseng, as well as other Panax species. It is also a reported neurotoxic agent found in Lathyrus sativus (grass pea seed) and cycad seeds. A method was developed for quantitative determination of the non-protein amino acid, dencichine, in plant samples of P. notoginseng and the adventitious roots directly from the explants of P. notoginseng after derivatization with ethyl chloroformate (ECF) by gas chromatography-mass spectrometry (GC-MS). l-2-chlorophenylalanine was used as an internal standard. Calibration curves were linear (r(2)=0.9988, n=6) in the range of 10-800 microg/ml for dencichine. Limit of detection and quantification for dencichine were 0.5 microg/ml and 2 microg/ml, respectively. This rapid and specific method may be applied to the quantification of dencichine in complex medicinal plants and their products.

[Application of metabolomics in research of plant metabolites].

Metabolomics, a branch of systems biology, has gained extensive attention and profound achievements in the plant. Although plant metabolomics is to be explored, it has been one of the most effective methods to study the physiological and biochemical process and gene modification in pattern plants. We herein summarized the concept, development, and application of metabolomics and prospected the potentials in the metabolite profiling for plant. Metabolomics provides an omics' methodology to elucidate the whole biological process, identify and quantify the complex components in the plant. A number of metabolites present in the plant are active components of traditional Chinese medicine, and these bioactive components are influenced by the multi-factors such as environment, species, and processing methods etc. Therefore, it is of great importance to analyze a wide spectrum of compositions with diverse chemical characteristics and varied concentration, which is the foundation to quality control, allowing the elucidation of the pharmacological effectiveness, and further exploiting of traditional Chinese medicine.

A new silymarin preparation based on solid dispersion technique.

Silymarin is a hepatoprotective agent that is poorly soluble in water. The present study describes a new preparation of solid dispersions in the form of "dripping pills" designed to enhance solubility. Dripping pills of silymarin were prepared at a 1:4 ratio by the traditional fusion method with the use of a mixture of silymarin and polyethylene glycol 6000 (PEG 6000). The prepared dripping pills were spherical and 3 to 4 mm in diameter, with an average weight of 30 mg per pill and with each pill containing 5 mg of silymarin. The dissolution rates of silymarin in dripping pill and of 3 other silymarin preparations, including Yiganling Film-Coating Tablet, Yiganling Sugar-Coating Tablet, and Legalon Capsule, were determined in pH 1.2 medium. The dissolution rate (T50) of the silymarin dripping pill was found to be significantly higher (by a factor of 7.5-11) than those of the other 3 preparations.