A Pyrazolate Osmium(VI) Nitride Exhibits Anticancer Activity through Modulating Protein Homeostasis in HepG2 Cells.

Interest in the third-row transition metal osmium and its compounds as potential anticancer agents has grown in recent years. Here, we synthesized the osmium(VI) nitrido complex Na[OsVI(N)(tpm)2] (tpm = [5-(Thien-2-yl)-1H-pyrazol-3-yl]methanol), which exhibited a greater inhibitory effect on the cell viabilities of the cervical, ovarian, and breast cancer cell lines compared with cisplatin. Proteomics analysis revealed that Na[OsVI(N)(tpm)2] modulates the expression of protein-transportation-associated, DNA-metabolism-associated, and oxidative-stress-associated proteins in HepG2 cells. Perturbation of protein expression activity by the complex in cancer cells affects the functions of the mitochondria, resulting in high levels of cellular oxidative stress and low rates of cell survival. Moreover, it caused G2/M phase cell cycle arrest and caspase-mediated apoptosis of HepG2 cells. This study reveals a new high-valent osmium complex as an anticancer agent candidate modulating protein homeostasis.

Osmium(VI) nitride triggers mitochondria-induced oncosis and apoptosis.

We report a new osmium(VI) nitrido complex bearing a nonplanar tetradentate ligand with potent anticancer activity. This complex causes mitochondrial damage, which induces liver cancer cell death via oncosis and apoptosis. This is the first osmium-based anticancer candidate that induces oncosis.

A cytotoxic nitrido-osmium(VI) complex induces caspase-mediated apoptosis in HepG2 cancer cells.

The osmium(vi) nitrido complex [OsVI(N)(sap)(py)Cl] is a potential anti-cancer drug with promising in vitro antiproliferative activities toward a panel of cancer cell lines, including cisplatin-resistant cells (IC50 values of 2.8-13.8 µM). This drug targets DNA and changes its conformation via covalent binding and insertion. In vitro studies indicate that the drug induces HepG2 cells G2/M phase arrest, disrupts the mitochondrial membrane potential and causes caspase-mediated apoptosis. Further in vivo studies using HepG2-bearing nude mice reveal that this drug not only shows good antitumor efficacy of inhibiting tumor growth, but also does not show the side effect of weight loss.

Geographical origin traceability of Cabernet Sauvignon wines based on Infrared fingerprint technology combined with chemometrics.

Mid-infrared (MIR) and near-infrared (NIR) spectroscopy combined with chemometrics were explored to classify Cabernet Sauvignon wines from different countries (Australia, Chile and China). Commercial wines (n = 540) were scanned in transmission mode using MIR and NIR, and their characteristic fingerprint bands were extracted at 1750-1000 cm-1 and 4555-4353 cm-1. Through the identification system of Tri-step infrared spectroscopy, the correlation between macroscopic chemical fingerprints and geographical regions was explored more deeply. Furthermore, Principal component analysis (PCA), soft independent modelling of class analogy (SIMCA) and discriminant analysis (DA) based on MIR and NIR spectra were used to visualize or discriminate differences between samples and to realize geographical origin traceability of Cabernet Sauvignon wines. Through "external test set (n = 157)" validation, SIMCA models correctly classified 97%, 97% and 92% of Australian, Chilean and Chinese Cabernet Sauvignon wines, while the DA models correctly classified 86%, 85% and 77%, respectively. Based on unique digital fingerprints of spectroscopy (FT-MIR and FT-NIR) associated with chemometrics, geographical origin traceability was achieved in a more comprehensive, effective and rapid manner. The developed database models based on IR fingerprint spectroscopy with chemometrics could provide scientific basis and reference for geographical origin traceability of Cabernet Sauvignon wines (Australia, Chile and China).

[System of simultaneous extraction and cleaning of 99 veterinary drugs in food].

We established a system for the simultaneous extraction and cleaning of 99 veterinary drugs from food by ultra-performance liquid chromatography coupled with quadrupole electrostatic field-orbital trap-high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). The system is based on the supported liquid-liquid extraction technology through one-off pretreatment, which covers the common eight categories of physical and chemical properties, and the extraction and purification of 99 different veterinary drug residues. At the same time, the one-step multi residue screening of 99 veterinary drug residues has been realized using HRMS. The results showed that the method was suitable for extraction from milk, pork, and fish. The 99 veterinary drugs showed a good linear relationship in the concentration range of 0.001-0.1 µg/mL, the limit of quantification was 0.5-20.0 µg/kg, the average recoveries were 60%-120%, and the relative standard deviations (RSDs) were not more than 15%. The pre-processing and instrumental analysis of this method have high detection efficiency, strong maneuver ability, and strong compatibility with different physical and chemical compounds. Moreover, the detection limit can meet all the objects of the test and greatly reduce the detection cost.

A Novel Zebrafish (Danio rerio) Assay for Assessing Musk Ambrette-Induced Toxicity.

Musk ambrette (4-tert-butyl-3-methoxy-2,6-dinitrotoluene) is a nitro musk, a cheap substitute for natural musk and a potential environmental pollutant based on its persistence, accumulation in human organisms. We investigated the acute toxicity of musk ambrette using wild-type AB and transgenic Tg(fli1a:EGFP)y1 zebrafish. Different concentrations were delivered to zebrafish by direct soaking from 6 to 72 h post-fertilization (hpf). The LC50 of musk ambrette was 76.4 µg mL-1. As musk ambrette concentration increased, zebrafish embryos showed developmental delays (50 µg mL-1, 22 hpf), pericardial edema (5 µg mL-1, 48 hpf), circulatory disturbances, curved body axis (1 µg mL-1, 72 hpf) and death (100 µg mL-1, 22 hpf). Target organ toxicity was evaluated by a zebrafish angiogenesis model. Musk ambrette induced cardiovascular morphological changes, vessel permeability variation, angiogenic changes and cardiotoxicity (10 µg mL-1, 48 hpf). The disappearance of caudal vein plexus confirmed the vascular development toxicity. Musk ambrette negatively affects early life-stage survival and demonstrates various toxicities in zebrafish.

Rapid identification of pearl powder from Hyriopsis cumingii by Tri-step infrared spectroscopy combined with computer vision technology.

Pearl powder, an important raw material in cosmetics and Chinese patent medicines, is commonly uneven in quality and frequently adulterated with low-cost shell powder in the market. The aim of this study is to establish an adequate approach based on Tri-step infrared spectroscopy with enhancing resolution combined with chemometrics for qualitative identification of pearl powder originated from three different quality grades of pearls and quantitative prediction of the proportions of shell powder adulterated in pearl powder. Additionally, computer vision technology (E-eyes) can investigate the color difference among different pearl powders and make it traceable to the pearl quality trait-visual color categories. Though the different grades of pearl powder or adulterated pearl powder have almost identical IR spectra, SD-IR peak intensity at about 861cm-1 (v2 band) exhibited regular enhancement with the increasing quality grade of pearls, while the 1082cm-1 (v1 band), 712cm-1 and 699cm-1 (v4 band) were just the reverse. Contrastly, only the peak intensity at 862cm-1 was enhanced regularly with the increasing concentration of shell powder. Thus, the bands in the ranges of (1550-1350cm-1, 730-680cm-1) and (830-880cm-1, 690-725cm-1) could be exclusive ranges to discriminate three distinct pearl powders and identify adulteration, respectively. For massive sample analysis, a qualitative classification model and a quantitative prediction model based on IR spectra was established successfully by principal component analysis (PCA) and partial least squares (PLS), respectively. The developed method demonstrated great potential for pearl powder quality control and authenticity identification in a direct, holistic manner.

[Rapid screening of three halogenated salicylanilines in cosmetics using ambient ionization coupled with ion mobility spectrometry].

OBJECTIVE: A rapid analytical method had been developed for the screening of three halogenated salicylanilines in cosmetics using ambition ionization coupled with ion mobility spectrometry. METHODS: Without time-consuming pretreatment, cosmetic samples of lotion, lipstick and powder were added or smudged onto a Grade 3MM chromatography paper for paper spray ionization, cosmetic samples of cream and shampoo were simply loaded with a metal wire probe for extraction nano-electrospray. RESULTS: The limits of detection( LODs) for the three halogenated salicylanilides were 50 µg/kg. The instrumental analysis time for a single ion mobility spectrometry run was less than 15 ms. The suspected positive samples were subjected to further verification by high-performance liquid chromatography coupled with tandem mass spectrometry. CONCLUSION: The method is simple, rapid, and with low cost and is suitable for rapid, on-site screening of cosmetics.

The complete mitochondria genome of Chrysomya phaonis (Seguy, 1928) (Diptera: Calliphoridae).

Chrysomya phaonis (Seguy, 1928) is one of the blowflies of great medical and forensic importance. In this paper, we report that the entire genome of mitochondrial DNA of C. phaonis is 15,831 bp in length, which consists of 39 genes including13 protein-coding genes, 24 tRNA genes, 2 mitochondrial ribosomal RNA genes, and a 992 bp non-coding A + T-rich region. The overall base compositions of A, G, C, and T are 38.79%, 9.75%, 14.15%, and 37.31%, respectively. We provide the first complete mitochondrial genome of C. phaonis, and should provide useful information for phylogenetic and species identification for C. phaonis.

Determination of a metabolite of nifursol in foodstuffs of animal origin by liquid-liquid extraction and liquid chromatography with tandem mass spectrometry.

An analytical method has been developed for the detection of a metabolite of nifursol, 3,5-dinitrosalicylic acid hydrazide, in foodstuffs of animal origin (chicken liver, pork liver, lobster, shrimp, eel, sausage, and honey). The method combines liquid chromatography and tandem mass spectrometry with liquid-liquid extraction. Samples were hydrolyzed with hydrochloric acid and derivatized with 2-nitrobenzaldehyde at 37°C for 16 h. The solutions of derivatives were adjusted to pH 7.0-7.5, and the metabolite was extracted with ethyl acetate. 3,5-Dinitrosalicylic acid hydrazide determination was performed in the negative electrospray ionization method. Both isotope-labeled internal standard and matrix-matched calibration solutions were used to correct the matrix effects. Limits of quantification were 0.5 µg/kg for all samples. The average recoveries, measured at three concentration levels (0.5, 2.0, and 10 µg/kg) were in the range of 75.8-108.4% with relative standard deviations below 9.8%. The developed method exhibits a high sensitivity and selectivity for the routine determination and confirmation of the presence of a metabolite of nifursol in foodstuffs of animal origin.

Determination of Six Pyrazole Fungicides in Grape Wine by Solid-Phase Extraction and Gas Chromatography-Tandem Mass Spectrometry.

A gas chromatography-tandem mass spectrometry (GC-MS/MS) method was developed for the first simultaneous identification and quantification of six pyrazole fungicides (furametpyr, rabenzazole, fluxapyroxad, penflufen, bixafen, and isopyrazam) in grape wine samples. The grape wine samples were first diluted with water, then purified by solid-phase extraction, and finally examined by GC-MS/MS in multiple reaction monitoring (MRM) mode. Matrix-matched calibration curves were used to correct the matrix effects. The limits of quantification (LOQs), calculated as 10 times the standard deviation, were 0.2-0.8 µg kg(-1) for the six pyrazole fungicides. The average recoveries were in the range of 74.3-94.5%, with relative standard deviations (RSDs) below 5.8%, measured at three concentration levels. The proposed method is suitable for the simultaneous determination of six pyrazole fungicides in grape wine samples.

Determination of validamycin A in agricultural food samples by solid-phase extraction combined with liquid chromatography-atmospheric pressure chemical ionisation-tandem mass spectrometry.

For the first time, a rapid, sensitive and accurate liquid chromatography-atmospheric pressure chemical ionisation-tandem mass spectrometry (LC-APCI-MS/MS) method was developed for determination of validamycin A in agricultural food samples (rice, agaric, almond, cabbage, green onion, carrot, tomato, cucumber and spinach). The validamycin A residue was extracted with methanol-water (9/1, v/v) or methanol by vortex, and a HLB solid-phase extraction cartridge was used for cleaning up the extracts. LC-APCI-MS/MS data acquisition was carried out in multiple reaction monitoring (MRM) mode. A series of matrix-matched calibration solutions ranging from 2.5 to 50ngmL(-1) were used to record calibration curve. The limit of quantification (LOQ) was 10µgkg(-1). The average recoveries, measured at three concentrations levels (10.0, 50.0, 100.0µgkg(-1)) were in the range 83.5-109.6%. The proposed method offers the best sensitivity and specificity for the routine analysis of validamycin A in agricultural food samples.

[Determination of metabolite residues of nitrofuran antibiotics in aquatic products by liquid chromatography-tandem mass spectrometry].

A method was developed for simultaneous qualitative and quantitative analysis of five metabolites of nitrofuran antibiotics, including 3-amino-2-oxazolidinone (AOZ), 5-morpholino-methyl-3-amino-2-oxazolidinone ( AMOZ ), semicarbazide ( SEM ), 1-aminohydantoin (AHD) and 3, 5-dinitrosalicylic acid hydrazide (DNSH) in aquatic products by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The samples were hydrolyzed with HCl, and derivatized with 2-nitrobenzaldehyde at 37 degre C for 16 hours. The derivative solutions were adjusted to pH 7.0 -7. 5, and the analytes were extracted by ethyl acetate. The separation was based on Thermo Aquasil C18 column (150 mm x 4.6 mm, 3.01 micro m). The analytes were detected by tandem mass spectrometry with electrospray ionization source with multiple reaction monitoring (MRM) mode. The developed method showed good linear correlation between the peak area ratios of the analyte and the internal standard and the concentration of the analyte with the correlation coefficients all above 0. 99 over the dynamic range of 0.5 - 10 micro g/kg. The limits of quantitation (LOQs) of AOZ, AMOZ, SEM, AHD and DNSH were 0.5 micro g/kg. The average recoveries of all the compounds at four spiked levels of 0.5, 1.0, 2.0 and 4. 0 micro g/kg ranged from 81.3% to 100.5% with the RSDs between 3.4% and 10.0% (n =6). The method is proved to be fast and effective for simultaneous qualitative and quantitative analysis of the metabolites of the nitrofuran antibiotics in aquatic products.

High human bocavirus viral load is associated with disease severity in children under five years of age.

Human bocavirus (HBoV) is a parvovirus and detected worldwide in lower respiratory tract infections (LRTIs), but its pathogenic role in respiratory illness is still debatable due to high incidence of co-infection with other respiratory viruses. To determine the prevalence of HBoV infection in patients with LRTI in Shanghai and its correlation with disease severity, we performed a 3-year prospective study of HBoV in healthy controls, outpatients and inpatients under five years of age with X-ray diagnosed LRTIs. Nasopharyngeal aspirates were tested by PCR for common respiratory viruses and by real time PCR for HBoV subtypes 1-4. Nasopharyngeal swabs from healthy controls and serum samples and stools from inpatients were also tested for HBoV1-4 by real time PCR. Viral loads were determined by quantitative real time PCR in all HBoV positive samples. HBoV1 was detected in 7.0% of inpatients, with annual rates of 5.1%, 8.0% and 4.8% in 2010, 2011 and 2012, respectively. Respiratory syncytial virus (RSV) subtype A was the most frequent co-infection detected; HBoV1 and RSVA appeared to co-circulate with similar seasonal variations. High HBoV viral loads (>10(6) copies/ml) were significantly more frequent in inpatients and outpatients than in healthy controls. There was a direct correlation of high viral load with increasing disease severity in patients co-infected with HBoV1 and at least one other respiratory virus. In summary, our data suggest that HBoV1 can cause LRTIs, but symptomatic HBoV infection is only observed in the context of high viral load.

[Identification of irradiated abalone by ESR spectroscopy].

OBJECTIVE: To establish an analytical method for the detection and identification of irradiated abalone by electron spin resonance spectroscopy. METHODS: Electron spin resonance (ESR) was used to study the spectral characteristics of abalone and the characteristic peak for quantitation. RESULTS: There were obvious different ESR spectra between unirradiated and irradiated abalone. The g factor for unirradiated abalone was 2.0055-2.0060, the g1 and g2 factor for irradiated abalone were (2.0027 +/- 0.0001) and (1.9994 +/- 0.0001), respectively. The ESR signal intensity of characteristic peak was positively correlated with absorbed dose in the range of 0.5 - 10 kGy, left peak was the characteristic peak for quantitation and the detection limit was < or = 0.5 kGy. It was difficult to quantitate when the absorbed dose was over 10 kGy. ESR characteristic peak and g factor were able to qualitatively determine the irradiation of abalone. CONCLUSION: ESR spectroscopy is an effective method to determine whether the abalone being irradiated or not.

[Discriminating and quantifying potential adulteration in virgin olive oil by near infrared spectroscopy with BP-ANN and PLS].

In the present paper, the use of near infrared spectroscopy (NIR) as a rapid and cost-effective classification and quantification techniques for the authentication of virgin olive oil were preliminarily investigated. NIR spectra in the range of 12 000 - 3 700 cm(-1) were recorded for pure virgin olive oil and virgin olive oil samples adulterated with varying concentrations of sesame oil, soybean oil and sunflower oil (5%-50% adulterations in the weight of virgin olive oil). The spectral range from 12 000 to 5 390 cm(-1) was adopted to set up an analysis model. In order to handle these data efficiently, after pretreatment, firstly, principal component analysis (PCA) was used to compress thousands of spectral data into several variables and to describe the body of the spectra, and the analysis suggested that the cumulate reliabilities of the first six components was more than 99.999%. Then ANN-BP was chosen as further research method. The six components were secondly applied as ANN-BP inputs. The experiment took a total of 100 samples as original model examples and left 52 samples as unknown samples to predict. Finally, the results showed that the 52 test samples were discriminated accurately. And the calibration models of quantitative analysis were built using partial-least-square (PLS). The R values for PLS model are 98.77, 99.37 and 99.44 for sesame oil, soybean oil and sunflower oil respectively, the root mean standard errors of cross validation (RMSECV) are 1.3, 1.1 and 1.04 respectively. Overall, the near infrared spectroscopic method in the present paper played a good role in the discrimination and quantification, and offered a new approach to the rapid discrimination of pure and adulterated virgin olive oil.

[A method of perfluorinated organic compounds in drinking water determinated by HPLC-MS/MS].

OBJECTIVE: To establish a method of perfluorinated organic compounds (POCs) in drinking water determinated by HPLC-MS/MS. METHODS: The contents of selected POCs (i. e., perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA) and perfluorodecanoate (PFDA)) in water samples were extracted by the C18 solid-phase extraction (SPE). The extraction samples were then identified and quantitated by high performance liquid chromatography-negative electrospray mass spectrometry (HPLC-ESI-MS/MS). RESULTS: Linear calibration curves were obtained at the concentration ranges from 50 pg/ml to 1000 pg/ml (the correlation coefficients were above 0.99). The average recoveries for PFOA, PFOS and PFDA spiked in water ranged from 65 to 111% and their relative standard were between 3.6% and 14.6%. The limit of quantification (LOQ) of the method was 8 pg/g. CONCLUSION: The method for determination of POCs in drinking water could be simple, sensitive and accurate.

[Determination of endosulfans and its metabolite in fishery foods using gas chromatography-mass spectrometry with negative ion chemical ionization].

Endosulfan is one of the organochliorine pesticides and contains two stereoisomers, alpha-endosulfan and beta-endosulfan. Endosulfan sulfate is the oxide of endosulfan with toxicity comparable to endosulfan. The method was developed for the analysis of alpha-endosulfan, beta-endosulfan and endosulfan sulfate in fishery foods. Since the maximal residue level is set at about 4 - 6 microg/kg in many countries, the acceptable method must be sensitive and accurate. The conditions were systematically optimized and external standard method was used for the quantification. The sample was extracted by acetone and hexane (1 : 1, v/v), removed fat by transfering to acetonitrile and cold, and cleaned up with an Envi-Carb column and an LC-Alumina-N column, and analyzed by gas chromatography-mass spectrometry negative ion chemical ionization with selective ion mode. The selective ions of endosulfans and endosulfan sulfate were m/z 406, 408, 404, 372 and m/z 386, 388, 384, 422, respectively. The lowest quantitative limited level of every compound was 0. 5 microg/kg, and the recoveries were 83% - 105%, and the relative standard deviations were less than 13%. The method is efficient, accurate and sensitive to meet the requirements for determining endosulfans in fishery foods.

[Study of normal reference values for bone mineral contents in children and adolescents in Beijing].

OBJECTIVE: The paper aims to provide reference values of bone mineral content for school-age children and adolescents in Beijing. METHODS: Normal values for total body bone mineral content (TBMC) were derived from measurements on 1025 children and adolescents aged from 6 to 18 years in Beijing city, and bone mineral content (BMC) values for selected regions of interest were also presented, including head, chest, midriff, pelvis, legs and arms. Both BMC and body mineral density were determined by dual-energy x-ray absorptiometry (DEXA). RESULTS: The results revealed that bone mineralization increased gradually in early childhood and accelerated during adolescence and there were significant age and gender effects on all regions of interest as well as total body. TBMC in 18 year group was about 3 and 2.7 times more respectively for male and female than that in 6 year group, and female's increasing trend flat at the age of 14, while male TBMC keep growing until 17. Legs contribute much more to the increase of TBMC than other regions. Compared with white-origin children, the subjects were bestowed with more TBMC but less TBMD. CONCLUSION: BMC depends on sex, age, region and race. The normal values can be used to the assessment of bone status for children and adolescences to serve clinical as well as research purposes.

[Association of Pro12Ala mutation in peroxisome proliferator-activated receptor gamma 2 with obesity and diabetes in Chinese population].

OBJECTIVE: To investigate peroxisome proliferator-activated receptors gamma 2 (PPAR gamma 2) polymorphism and its association with obesity or diabetes in Chinese population. METHODS: Normal weight group(91) and overweight and obese group(140) were included in this study, according to BMI. 103 subjects had normal glucose tolerance, 50 had impaired glucose tolerance, and 78 had diabetes mellitus, according to WHO Criteria for Diagnosis of Diabetes Mellitus. We evaluated these subjects for the Pro12Ala mutation in the PPAR gene using PCR-restriction fragment length polymorphism. RESULTS: Allele frequencies of Pro12Ala mutation of PPAR gamma 2 in overweight and obese subjects (qA = 3.93%) were not significantly different from those in normal weight group (qA = 3.85%, P > 0.05). Both in overweight and obese group and normal weight group, the genotype of PA(AA) had lower plasma resistin as compared with the genotype of PP(P < 0.05). Allele frequencies of Pro12Ala were not different among Chinese subjects with normal glucose tolerance (qA = 3.88%), those with impaired glucose tolerance (qA = 4.00%), and those with diabetes mellitus (qA = 3.85%, P > 0.05). In these three groups, the genotype of PA(AA) had lower plasma resistin as compared with the genotype of PP, only in impaired glucose tolerance group the difference was not significant (P > 0.05). CONCLUSION: The Pro12Ala mutation in PPAR gamma 2 was not associated with either diabetes or obesity. The PPAR gamma 2 Ala allele may be involved in the expression of resistin and insulin sensitivity.