Dissection of flag leaf metabolic shifts and their relationship with those occurring simultaneously in developing seed by application of non-targeted metabolomics.

Rice flag leaves are major source organs providing more than half of the nutrition needed for rice seed development. The dynamic metabolic changes in rice flag leaves and the detailed metabolic relationship between source and sink organs in rice, however, remain largely unknown. In this study, the metabolic changes of flag leaves in two japonica and two indica rice cultivars were investigated using non-targeted metabolomics approach. Principal component analysis (PCA) revealed that flag leaf metabolomes varied significantly depending on both species and developmental stage. Only a few of the metabolites in flag leaves displayed the same change pattern across the four tested cultivars along the process of seed development. Further association analysis found that levels of 45 metabolites in seeds that are associated with human nutrition and health correlated significantly with their levels in flag leaves. Comparison of metabolomics of flag leaves and seeds revealed that some flavonoids were specific or much higher in flag leaves while some lipid metabolites such as phospholipids were much higher in seeds. This reflected not only the function of the tissue specific metabolism but also the different physiological properties and metabolic adaptive features of these two tissues.

Relationship between smoking status and muscle strength in the United States older adults / 한국역학회지

OBJECTIVES@#Muscle strength in older adults is associated with greater physical ability. Identifying interventions to maintain muscle strength can therefore improve quality of life. The purpose of this study was to evaluate whether current or former smoking status is associated with a decrease in muscle strength in older adults. @*METHODS@#Data from the Health and Retirement Study from 2012-2014 were analyzed with regard to maximum dominant hand grip strength, maximum overall hand grip strength, and smoking status (current, former, or never). Unadjusted linear regression was conducted. Other factors known to be related to strength were included in the adjusted linear regression analyses. @*RESULTS@#For maximum grip strength, the regression coefficient was 4.91 for current smoking (standard error [SE], 0.58; p<0.001), 3.58 for former smoking (SE, 0.43; p<0.001), and 28.12 for never smoking (SE, 0.34). Fully adjusted linear regression on the relationship between dominant hand grip strength and smoking did not yield a significant result. The factors significantly associated with dominant hand grip strength were male sex, younger age, a race/ethnicity of non-Hispanic White or non-Hispanic Black, higher income, morbidity of ≤1 condition, no pain, and moderate or vigorous exercise more than once a week. @*CONCLUSIONS@#Muscle strength in older adults was not associated with smoking status in the adjusted analysis.

Relationship between smoking status and muscle strength in the United States older adults / 한국역학회지

OBJECTIVES@#Muscle strength in older adults is associated with greater physical ability. Identifying interventions to maintain muscle strength can therefore improve quality of life. The purpose of this study was to evaluate whether current or former smoking status is associated with a decrease in muscle strength in older adults. @*METHODS@#Data from the Health and Retirement Study from 2012-2014 were analyzed with regard to maximum dominant hand grip strength, maximum overall hand grip strength, and smoking status (current, former, or never). Unadjusted linear regression was conducted. Other factors known to be related to strength were included in the adjusted linear regression analyses. @*RESULTS@#For maximum grip strength, the regression coefficient was 4.91 for current smoking (standard error [SE], 0.58; p<0.001), 3.58 for former smoking (SE, 0.43; p<0.001), and 28.12 for never smoking (SE, 0.34). Fully adjusted linear regression on the relationship between dominant hand grip strength and smoking did not yield a significant result. The factors significantly associated with dominant hand grip strength were male sex, younger age, a race/ethnicity of non-Hispanic White or non-Hispanic Black, higher income, morbidity of ≤1 condition, no pain, and moderate or vigorous exercise more than once a week. @*CONCLUSIONS@#Muscle strength in older adults was not associated with smoking status in the adjusted analysis.

Quality Evaluation and Identification of Phyllanthi Fructus at Different Storage Years Based on HPLC-UV and FT-NIR / 中国实验方剂学杂志

Objective: To explore the change rules of active ingredients in Phyllanthi Fructus of different storage years,in order to provide theory basis for storage. Method: Seven Phyllanthi Fruatus samples of different storage years were collected. HPLC-UV detection method was established to determine the contents of gallic acid,corilagin,chebulagic acid,ellagic acid and quercetin. Samples were fingerprinted by FT-NIR and identified by PLS-DA model. Result: Gallic acid,which was the bioactive marker in Chinese Pharmacopoeia,had the highest content. It was followed by ellagic acid and chebulagic acid,and corilagin and quercetin had the least content. The components had significant differences between samples of different storage years (P-1 respectively. The contents of chebulagic acid,corilagin and ellagic acid reached a maximum at 4 years of storage,which were 18.85,7.97,21.46 mg·g-1,respectively. FT-NIR data was optimized by MSC+SG (second derivative, the window parameter as 11,and the polynomial order as 3). The classification accuracy was 84.5%. Spectral data reduced to several important potential variables,and was fused with 5 active components based on minimum cross-validation root mean square error,and the classification accuracy increased to 98.8%. Conclusion: The analysis of PLS-DA by HPLC-UV and FT-NIR could effectively explain the accumulation characteristics of active components in Phyllanthi Fruatus. According to the data fusion strategy,PLS-DA model could distinguish samples of different qualities. The results provide a scientific basis for the quality evaluation and identification of Phyllanthi Fruatus.

Diagnostic value of plasma tryptophan and symmetric dimethylarginine levels for acute kidney injury among tacrolimus-treated kidney transplant patients by targeted metabolomics analysis.

Few literatures have evaluated the exact role of metabolomics in the identification process of potential biomarkers for acute kidney injury among the patients receiving renal transplantation. On top of this, the success of metabolomics in biomarker translation seems to lie in the robust quantitative method. As such, a single-center retrospective observational study was conducted enrolling 42 patients underwent renal transplantation with/without acute kidney injury, as well as 24 healthy volunteers, in Shanghai Changzheng Hospital. Plasma amino acid metabolic patterns for the participants were investigated by targeted UHPLC-MS/MS metabolic profiling. The most significant changes of the explored metabolites were related to the disturbance of tryptophan metabolism and arginine metabolism. Abnormal circulating tryptophan and symmetric dimethylarginine were identified to be potential biomarkers of acute kidney injury, combination of which showed a higher area under receiver-operator curve value (AUC = 0.901), improved sensitivity (0.889) and specificity (0.831) compared with creatinine only. Overall, these results revealed that targeted metabolomics analysis would be a potent and promising strategy for identification and pre-validation of biomarkers of acute kidney injury in renal transplantation patients.

Occurrence and estrogenic activity of steroid hormones in Chinese streams: A nationwide study based on a combination of chemical and biological tools.

Steroid hormones (SHs) are continuously released into the aquatic environment through various pathways after being excreted by humans and animals, interfere with the normal function of the endocrine system and may affect the physiology and reproduction of exposed aquatic life. To conduct a nationwide investigation of the occurrence and biological effects of SHs in surface river/steam water in China, we quantitated 27 selected SHs in 217 surface water samples by solid-phase extraction (SPE) tandem LC-MS/MS and used a recombinant yeast estrogen assay to screen extracts of the water samples for estrogenic activities. SHs were commonly found in the surface water samples, and their levels were typically in the ng L-1 range. Estrone (E1) and estriol (E3) were normally present in several to dozens of times higher concentrations than estradiol (E2) and 17-a-Ethinylestradiol (EE2). The high concentrations (mean > 1 µg L-1) of SumSHs were primarily obtained in areas under extreme water stress, specifically the eastern coastal areas. Source apportionment based on the profiles of the target compounds indicated that 54.5% of the SHs in target samples came from freshly discharged untreated sewage. The estrogen equivalent (EEQ(bio)) values ranged from 0.01 to 40.27 ng L-1, and the calculated EEQ (EEQ(cal)) values were generally lower than the corresponding EEQ(bio) values for all samples. E2 was the main contributor to the estrogenicity among the three estrogens, with a contribution ratio of 82.8%. The risk quotient values of E2 were highest and ranged from 1.55 to 782.95, and 76.0% of the target surface samples displayed the greatest environmental risk. We concluded that the impacts of SHs on humans in Chinese surface waters should not be ignored and that certain actions should be taken to decrease the levels of SHs in source waters, especially measures targeting SHs in untreated wastewater from the vast rural areas.

Targeted metabolomic analysis of 33 amino acids and biogenic amines in human urine by ion-pairing HPLC-MS/MS: Biomarkers for tacrolimus nephrotoxicity after renal transplantation.

Calcineurin inhibitor nephrotoxicity, especially for the widely used tacrolimus, has become a major concern in post-transplant immunosuppression. Multiparametric amino acid metabolomics is useful for biomarker identification of tacrolimus nephrotoxicity, for which specific quantitative methods are highlighted as a premise. This article presents a targeted metabolomic assay to quantify 33 amino acids and biogenic amines in human urine by high-performance liquid chromatography coupled with tandem mass spectrometry. Chromatographic separation was carried out on an Agilent Zorbax SB-C18 column (3.0 × 150 mm, 5 µm) with addition of an ion-pairing agent in the mobile phase, and MS/MS detection was achieved in both the positive and negative multiple reaction monitoring modes. Good correlation coefficients (r2 > 0.98) were obtained for most analytes. Intra- and inter-day precision, stability, carryover and incurred sample reanalysis met with the acceptance criteria of the guidance of the US Food and Drug Administration. Analysis on urine from healthy volunteers and renal transplantation patients with tacrolimus nephrotoxicity confirmed symmetric dimethylarginine and serine as biomarkers for kidney injury, with AUC values of 0.95 and 0.81 in receiver operating characteristic analysis, respectively. Additionally, symmetric dimethylarginine exhibited a tight correlation with serum creatinine, and was therefore indicative of renal function. The targeted metabolomic assay was time and cost prohibitive for amino acid analysis in human urine, facilitating the biomarker identification of tacrolimus nephrotoxicity.

Photochemical Transformation of Nicotine in Wastewater Effluent.

Nicotine is a highly toxic tobacco alkaloid that is ubiquitous in wastewater effluent. For the first time, we report the identification of the products and the pathways for the photodegradation of nicotine in an effluent matrix under simulated solar irradiation. Nicotine was found to be degraded by triplet-state organic matter (3OM*), thus indicating that electron transfer is a preferred reaction mechanism. Using the multivariate statistical strategies orthogonal projection to latent structures discriminant analysis (OPLS-DA) and hierarchical clustering, 49 potential transformation products (TPs) of nicotine were successfully extracted from the water matrix via high-resolution ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS). Overall, 30 TPs, including 4 groups of nonseparated isomeric photo TPs, were identified with various levels of confidence based on the tandem mass spectrometry information on standard compounds and the isotope-labeling method (using rac-nicotine-2',3',3'-D3, rac-nicotine-13CD3, and rac-nicotine-D4) under air-saturated conditions. The pyrrolidine ring of nicotine was found to be the reactive site under sunlight irradiation. Pseudooxynicotine was the main primary TP from nicotine, with a maximum transformation ratio of 64%. Nicotinic acid, cotinine, 3'-hydroxycotinine, and myosmine were the final stable TPs after 72 h of solar irradiation, with yields of 13%, 3%, 5%, and 5%, respectively.

Long-term outcome of phase I/II prospective study of dose-escalated proton therapy for early-stage non-small cell lung cancer.

PURPOSE: The aim of this phase I/II study was to assess the long-term clinical benefits and toxicities of proton beam therapy for medically inoperable early-stage non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: From June 2006 to September 2011, 35 patients with medically inoperable T1N0M0 (central or superior location, 12 patients) or T2-3N0M0 (any location, 23 patients) NSCLC were treated with 87.5Gy at 2.5Gy/fraction of proton therapy. Toxicities were scored according to the Common Terminology Criteria for Adverse Events, version 4.0. RESULTS: The median follow-up time was 83.1months (95% CI: 69.2-97.1months). For all 35 patients, the 1, 3, and 5-year overall survival rates were 85.7%, 42.9%, and 28.1%, respectively. The 5-year local recurrence-free, regional recurrence-free, and distant metastasis-free survival rates were 85.0%, 89.2%, and 54.4%, respectively. Different T stages had no effect on local and regional recurrence (p=0.499, p=1.00). However, with the increase in T stages, the distant metastasis rate increased significantly (p=0.006). The most common adverse effects were dermatitis (grade 2, 51.4%; grade 3, 2.9%) and radiation pneumonitis (grade 2, 11.4%; grade 3, 2.9%). Other grade 2 toxicities included esophagitis (2.9%), rib fracture (2.9%), heart toxicities (5.7%), and chest wall pain (2.9%). CONCLUSIONS: According to our long-term follow-up data, proton therapy with ablative doses is well tolerated and effective in medically inoperable early-stage NSCLC. Systemic therapy should be considered to reduce the rate of distant metastasis in cases of T2 and T3 lesions.

Differential Expression of CD269 and CD317 Genes in Bone Marrow of Patients with Multiple Myeloma / 现代检验医学杂志

Objective To explore the differential expression of CD269 and CD317 in patients with multiple myeloma(MM). Methods Newly diagnosed samles from patients of MM(20 cases)and iron deficiency anemia(20 cases),40 cases in total (from 06/2015 to 08/2013,the Department of Hematology,Central Hospital of Zhuzhou City)were collected.Real-time quantitative PCR(RQ-PCR)tests were used to detect the relative expression of CD269 and CD317 in bone marrow sam-ples,and the results were statistically analyzed with clinical features.Results The relative expression levels of CD269 and CD317 in patients with multiple myeloma(4.418±4.568,4.327±2.876)were significantly higher than those in the control group(0.600±0.838,1.033±1.335),the difference was statistically significant(t=3.676,4.646,all P<0.05)respective-ly,while not related with the gender,age(P>0.05).There was no correlation between the expression of CD269 and CD317 (r=0.041,P=0.864),but positively correlated with the ratio of myeloma cells(r=0.495,P=0.026;r=0.533,P=0.016).Conclusion CD269 and CD317 were highly expressed in patients with multiple myeloma and may be involved in the pathogenesis of multiple myeloma.

Determination of illicit drugs in aqueous environmental samples by online solid-phase extraction coupled to liquid chromatography-tandem mass spectrometry.

In this study, a fully automated analytical method, based on online solid phase extraction coupled to liquid chromatography-triple quadrupole tandem mass spectrometry (online SPE-LC-MS/MS), has been developed and optimized for the quantification of 10 illicit drugs and metabolites in environmentally aqueous samples collected from China. The particular attention was devoted to minimize the matrix effects through a washing step, which washed out the interferences effectively and helped to reduce the matrix effect significantly. The key advantages of the method are high sensitivity, selectivity and reliability of results, smaller sample manipulation, full automation, and fairly high throughput. The whole procedure was then successfully applied in the analysis of various surface water and wastewater effluents samples. Pseudoephedrine have been detected at trace levels (several tens ng L(-1) or less), while MDA, MDMA, benzoylecgonine and methadone were below the LOQ in all samples. Caffeine, cotinine and paraxanthine, which may be derived from medicines and foods, were detected with the highest frequencies and concentrations.

Identification of Conserved and Diverse Metabolic Shifts during Rice Grain Development.

Seed development dedicates to reserve synthesis and accumulation and uncovering its genetic and biochemical mechanisms has been a major research focus. Although proteomic and transcriptomic analyses revealed dynamic changes of genes and enzymes involved, the information regarding concomitant metabolic changes is missing. Here we investigated the dynamic metabolic changes along the rice grain development of two japonica and two indica cultivars using non-targeted metabolomics approach, in which we successfully identified 214 metabolites. Statistical analyses revealed both cultivar and developmental stage dependent metabolic changes in rice grains. Generally, the stage specific metabolic kinetics corresponded well to the physiological status of the developing grains, and metabolic changes in developing rice grain are similar to those of dicot Arabidopsis and tomato at reserve accumulation stage but are different from those of dicots at seed desiccation stage. The remarkable difference in metabolite abundances between japonica and indica rice grain was observed at the reserve accumulation stage. Metabolite-metabolite correlation analysis uncovered potential new pathways for several metabolites. Taken together, this study uncovered both conserved and diverse development associated metabolic kinetics of rice grains, which facilitates further study to explore fundamental questions regarding the evolution of seed metabolic capabilities as well as their potential applications in crop improvement.

Characterization and evaluation of two-dimensional microfluidic chip-HPLC coupled to tandem mass spectrometry for quantitative analysis of 7-aminoflunitrazepam in human urine.

Microfluidic chip-based high-performance-liquid-chromatography coupled to mass spectrometry (chip-HPLC-MS) has been widely used in proteomic research due to its enhanced sensitivity. We employed a chip-HPLC-MS system for determining small molecules such as drug metabolites in biological fluids. This chip-HPLC-MS system integrates a microfluidic switch, a 2-dimensional column design including an enrichment column (160 nL) for sample pre-concentration and an analytical column for chromatographic separation, as well as a nanospray emitter on a single polyimide chip. In this study, a relatively large sample volume (500 nL) was injected into the enrichment column for pre-concentration and an additional 4 µL of the initial mobile phase was applied to remove un-retained components from the sample matrix prior to chromatographic separation. The 2-dimensional column design provides the advantages of online sample concentration and reducing matrix influence on MS detection. 7-Aminoflunitrazepam (7-aminoFM2), a major metabolite of flunitrazepam (FM2), was determined in urine samples using the integrated chip-HPLC-MS system. The linear range was 0.1-10 ng mL(-1) and the method detection limit (signal-to-noise ratio of 3) was 0.05 ng mL(-1) for 7-aminoFM2. After consecutive liquid-liquid extraction (LLE) and solid-phase extraction (SPE), the chip-HPLC-MS exhibited high correlation between 7-aminoFM2 spiked Milli-Q water and 7-aminoFM2 spiked urine samples. This system also showed good precision (n = 5) and recovery for spiked urine samples at the levels of 0.1, 1.0, and 10 ng mL(-1). Intra-day and inter-day precision were 2.0-7.1% and 4.3-6.0%, respectively. Clinical urine samples were also analyzed by this chip-HPLC-MS system and acceptable relative differences (-1.3 to -13.0%) compared with the results using a GC-MC method were determined. Due to its high sensitivity and ease of operation, the chip-HPLC-MS system can be utilized for the determination of small molecules such as drug metabolites and neurotransmitters in biological fluids for clinical diagnosis.

Effects of emulsified isoflurane preconditioning on myocardial NF-κB activity during ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To investigate the effects of emulsified isoflurane preconditioning on myocardial NF-κB activity during ischemia-reperfusion(I/R)in rats.Methods Forty-eight healthy male SD rats weighing 230-280 g were randomly divided into 4 groups with 12 animals in each group: sham operation group(group S),I/R group,lipid emulsion + I/R group(group L)and emulsified isoflurane + I/R group(group EI).In group I/R,EI and L,myocardial I/R was produced by occlusion of left coronary anterior descending artery(LAD)for 30 min followed by 120 min reperfusion.In group L,30% lipid emulsion 4 ml·kg-1 ·h-1 was infused intravenously for 30 min before myocardial I/R.In group EI,emulsified isoflurane 4 ml· kg- 1 · h- 1 was infused intravenously for 30 min followed by 15 min washout before myocardial I/R.In group S and I/R,normal saline was given instead.Blood samples were taken from femoral artery at the end of 120 min reperfusion for determination of serum cTnI and IL-6 concentrations and CK-MB activity by ELISA.The rats were then killed and the myocardial tissues were taken for determination of NF-κB activity by Western blot and observation of the ultrastructure by electron microscopy.Results The NF-κB activity,serum cTnI and IL-6 concentrations and CK-MB activity were significantly higher in group I/R,EI and L than in group S(P ＜ 0.05 or 0.01),while lower in group EI than in group I/R(P ＜ 0.05).Microscopic examination showed that emulsified isoflurane significantly attenuated the histopathological changes in group EI.Conclusion Emulsified isoflurane pretreatment can attenuate myocardial I/R injury through decreasing the NF-κB activity and inhibiting inflammatory response in rats.