Sodium Houttuyfonate Alleviates Monocrotaline-induced Pulmonary Hypertension by Regulating Orai1 and Orai2.

An increased intracellular Ca2+ concentration ([Ca2+]i) is a key trigger for pulmonary arterial smooth muscle cell (PASMC) proliferation and contributes greatly to pulmonary hypertension (PH). Extracellular Ca2+ influx via a store-operated Ca2+ channel, termed store-operated Ca2+ entry (SOCE), is a crucial mechanism for [Ca2+]i increase in PASMCs. Calcium release-activated calcium modulator (Orai) proteins, consisting of three members (Orai1-3), are the main components of the store-operated Ca2+ channel. Sodium houttuyfonate (SH) is a product of the addition reaction of sodium bisulfite and houttuynin and has antibacterial, antiinflammatory, and other properties. In this study, we assessed the contributions of Orai proteins to monocrotaline (MCT)-enhanced SOCE, [Ca2+]i, and cell proliferation in PASMCs and determined the effect of SH on MCT-PH and the underlying mechanism, focusing on Orai proteins, SOCE, and [Ca2+]i in PASMCs. Our results showed that: 1) Orai1 and Orai2 were selectively upregulated in the distal pulmonary arteries and the PASMCs of MCT-PH rats; 2) knockdown of Orai1 or Orai2 reduced SOCE, [Ca2+]i, and cell proliferation without affecting their expression in PASMCs in MCT-PH rats; 3) SH significantly normalized the characteristic parameters in a dose-dependent manner in the MCT-PH rat model; and 4) SH decreased MCT-enhanced SOCE, [Ca2+]i, and PASMC proliferation via Orai1 or Orai2. These results indicate that SH likely exerts its protective role in MCT-PH by inhibiting the Orai1,2-SOCE-[Ca2+]i signaling pathway.

Magnetic Solid-Phase Extraction Based on Magnetic Sulfonated Reduced Graphene Oxide for HPLC-MS/MS Analysis of Illegal Basic Dyes in Foods.

In this study, a magnetic solid-phase extraction (MSPE) method coupled with High-Performance Liquid Chromatography Mass Spectrometry (HPLC-MS/MS) for the determination of illegal basic dyes in food samples was developed and validated. This method was based on Magnetic sulfonated reduced graphene oxide (M-S-RGO), which was sensitive and selective to analytes with structure of multiaromatic rings and negatively charged ions. Several factors affecting MSPE efficiency such as pH and adsorption time were optimized. Under the optimum conditions, the calibration curves exhibited good linearity, ranging from 5 to 60 µg/g with correlation coefficients >0.9950. The limits of detection of 16 basic dyes were in the range of 0.01-0.2 µg/L. The recoveries ranged from 70% to 110% with RSD% < 10%. The results indicate that M-S-RGO is an efficient and selective adsorbent for the extraction and cleanup of basic dyes. Due to the MSPE procedures, matrix effect and interference were eliminated in the analysis of HPLC-MS/MS without the matrix-matched standards. Thus, validation data showed that the proposed MSPE-HPLC-MS/MS method was rapid, efficient, selective, and sensitive for the determination of illegal basic dyes in foods.

Anti-Inflammatory and Anti-Aging Evaluation of Pigment-Protein Complex Extracted from Chlorella Pyrenoidosa.

Oxidative stress contributes to chronic inflammatory processes implicated in aging, referred to as "inflamm-aging." In this study, the potential anti-inflammatory and anti-aging effects of a pigment-protein complex (PPC) from Chlorella pyrenoidosa were investigated using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and D-galactose (D-gal)-induced aging in a murine model. Results indicated that PPC inhibits the production of the inflammatory cytokines TNF-α and IL-6, and the inflammatory mediator nitric oxide (NO) in LPS-stimulated RAW 264.7 cells. It also protected mice from D-gal induced informatory aging by increasing the activity of the antioxidant enzyme, such as superoxide dismutase (SOD), inhibiting D-gal-induced NF-κB upregulation, and increasing PPARs expression in the brain and gut. The findings indicated that PPC has favorable anti-inflammatory and anti-aging properties, and could be useful in the treatment of acute inflammation and senescence diseases.

Separation and Lipid Inhibition Effects of a Novel Decapeptide from Chlorella pyenoidose.

A novel lipid inhibition peptide Leu-Leu-Val-Val-Try-Pro-Trp-Thr-Gln-Arg (PP1) (MW 1274.53 Da) was obtained from Chlorella pyenoidose using enzymatic hydrolysis, gel filtration chromatography, and LC-MS/MS. Its lipid inhibition effects indicated that the synthetic peptide PP1 exhibits a good inhibitory effect against porcine pancreatic lipase (PL) (47.95%) at 200 µg/mL, which could be attributed to its hydrogen binding into catalytic sites of PL (Ser153, Asp177, and His 264) by docking analysis. Furthermore, in 3T3-L1 cells, the synthetic PP1 remarkedly decreased the accumulation of intracellular triacylglycerol (27.9%, 600 µg/mL), which carried a similar consequence as the positive drug simvastatin (24.1%, 10 µM). Western blot revealed that PP1 inhibited the lipid accumulation and fatty acid synthesis in 3T3-L1 adipocytes in two pathways, primarily: nonalcoholic fatty liver disease (NAFLD) pathway (C/EBPα, SREBP-1c, AMPKα) and AMPK signaling pathway (SREBP-1c, PPARγ, AMPKα). In short, these results support that PP1 can be used as a potential agent against obesity.

Simultaneous qualitative and quantitative determination of 104 fat-soluble synthetic dyes in foods using disperse solid-phase extraction and UHPLC-Q-Orbitrap HRMS analysis.

Public exposure to synthetic dyes through foods has attracted ongoing and serious attention. Here we developed and validated a simultaneous screening and quantitation method for the analysis of fat-soluble synthetic dyes that most frequently found in foods, using C18 d-SPE clean-up and UHPLC-Q-Orbitrap HRMS on Full-MS/dd-MS2 mode. During a single run, 104 dyes including 6 pairs of isomers were distinguished based on chromatographic separation and unique product ions. The method showed satisfactory linearity (R > 0.99), recoveries (61.3 %-118.8 %), precision (<20 %) and limit of quantification (0.05-0.5 mg/kg). For 98 % of test dyes, screening detection limits ranged from 2.5 to 25 µg/kg. The validated method was successfully applied to real commercial foodstuffs revealing the presence of two selected illegal dyes in three samples.

Qualitative screening and quantitative determination of multiclass water-soluble synthetic dyes in foodstuffs by liquid chromatography coupled to quadrupole Orbitrap mass spectrometry.

A LC-Q-Orbitrap HRMS analytical method for both qualitative screening and quantitative determination of 90 synthetic dyes including ten groups of isomers in foods has been established. An in-house synthetic dyes database and characteristic ions were also developed. Based on Q-Orbitrap HRMS, mass spectrum and fragmentation patterns of synthetic dyes were studied, which indicated that double charged ions were usually the main precursor ions. Matrix effects were successfully eliminated by the C18 d-SPE clean-up coupled with dilute and shoot approach with methanol-water (1:4, v/v) in 100-fold. For most of the compounds, mean recoveries were satisfactory between 70% and 120% with RSD < 20% at three spiked level in the range of 0.025-1.0 mg/kg. The screening detection limits ranged from 0.025 - 1.0 mg/kg. Method validation showed that the established method was efficient, rapid and high-throughput, which has been successfully applied to the monitoring of these water-soluble synthetic dyes in foods.

Primary anorectal malignant melanoma: a case report.

Anorectal melanoma (ARM) is rare and lethal. We report a case of a 48-year-old woman with 9 months of rectal swelling and bleeding. Physical examination revealed a mass about 5 × 6 cm on the anterior wall of the rectum, 3 cm from the anal verge, and the patient underwent abdominoperineal resection (APR). After hematoxylin-eosin staining and immunohistochemical staining, it was considered an ARM, which is an aggressive disease with a poor survival. Immunohistochemical staining showed the tumor to be positive for S-100, Melan A, Ki67 proliferative index of 70%, and negative for HMB45. The melanoma had infiltrated the adventitia and metastasized to the (intestinal) 16/16 lymph nodes with cancerous nodule formation. There were multiple organs with metastasis (liver, spleen, pancreas, lung and subcutaneous soft tissue) three months after operation. Overall, pre-operative biopsy may be insufficient to make a definite diagnosis, and immunohistochemistry is necessary. Therefore, the gold standard treatment for ARM is oncological radical surgical resection.

Simultaneous determination of multiclass illegal dyes with different acidic-basic properties in foodstuffs by LC-MS/MS via polarity switching mode.

Simultaneous determination of multiclass illegal dyes possessing different chemical properties is difficult. By using LC-MS/MS via negative/positive ion switching mode, an efficient and fast multi-residual method for simultaneous determination of multiclass 52 illegal dyes with different acidic-basic properties in foodstuffs was developed and validated during one single run, including 23 fat-soluble neutral azo dyes, 8 acidic sulfonated azo dyes, 12 triphenylmethane basic dyes, three basic indole dyes, three xanthene dyes, one quinoline dye, and two anthraquinones dyes. The illegal dyes were extracted with methanol-acetonitrile and further purified with d-SPE procedure to reduce interference. Sample dilution with 100-fold was used for the elimination of matrix effects of the quantitation of LC-MS/MS analysis. Validation data showed the good recoveries in the range of 71.2-111.2%, with relative standard deviations less than 20%, suggesting the developed method is suitable for the identification and quantitation of multiclass illegal dyes at trace levels in foods.

[Investigation on a seasonal influenza accompanying with the first locale novel A/H1N1 influenza outbreak in China].

OBJECTIVE: To timely summarize past experience and to provide more pertinent reference for control and prevention in A/H1N1 cases in influenza season. METHODS: During May 25 to 31, 2009, 2 secondary community cases caused by a influenza A/H1N1 imported case. In the close contacts of 3 A/H1N1 cases, 14 had some aspirator symptoms onset, such as fever (> or = 37.5 degrees C), cough, sore throat and etc. Laboratory tests excluded the infection of A/H1N1 influenza. For throat swab test for the 14 cases, 7 were tested for seasonal influenza virus. A face-to-face or telephone interview was conducted by CDC staff to collect information of 62 close contacts. RESULTS: Of 14 fever cases, there was no significant by differences by age[15-age group: 19.2% (5/26), over 25-age group: 25.0% (9/36); chi(2) = 0.287, P = 0.592]; by sex group [24.0% (6/25) for male and 21.6% (8/37) for female; chi(2) = 0.048, P = 0.826], by working units [dressing and design, photograph, saleroom and others, consumer group: 42.1% (8/19), 27.3% (3/11), 12.5% (2/16) and 6.3% (1/16); chi(2) = 7.653, P = 0.054], by dormitory style [dormitory style = 33.3% (4/12), non-dormitory style = 29.4% (10/34); chi(2) = 0.699, P = 0.403]. All the cases had fever (37.5 - 37.9 degrees C), no case had diarrhea. One in 3 A/H1N1 cases had diarrhea. All the 14 cases were negative result for A/H1N1 RNA. Six from 7 cases were positive for seasonal influenza test. CONCLUSION: This was a seasonal influenza outbreak happened in the close contacts of first confirmed A/H1N1 cases in community in mainland China. It showed that we should exclude the seasonal influenza in the investigation of A/H1N1 cases in the seasonal influenza period in some time. It is necessary to take effective measure to strengthen the control and prevention of seasonal influenza.

Pituitary involvement of langerhans cell histiocytosis in an adult unveiled by FDG PET/CT.

A 39-year-old woman with complains of polydipsia and dieresis was admitted to endocrinology service for evaluation and potential treatment of diabetes insipidus. During the hospitalization, the patient was found to have unexplained fever, for which an FDG PET/CT was performed to explore the source. The images revealed multiple regions of abnormal hypermetabolism throughout the body. Notably, there was an intense activity in the region of pituitary stalk. Pathological results after biopsy showed pituitary involvement of Langerhans cell histiocytosis.

[Epidemiological characteristic of first case of locally identified A/H1N1 secondary cases caused by imported source of infection in China].

OBJECTIVE: To study the first locally identifed A/H1N1 secondary cases outbreak in China. METHODS: Interview and field investigation were integrated to describe the whole process of transmission on each case and to illustrate the relationships between the onset of the disease and the retated factors. RESULTS: Two contact persons appearanced fever and whose throat swabs were tested positive to H1N1 viral nucleic acid. The two had a history of contact in a short distance with the initial imported case without any protective measure in the poor air ventilation. The patients clinical situation was slight. The incubation was between 37 hours and 57 hours. No other new case was found after intervention as isolation and antisepsis were taken. CONCLUSION: This event was proved to be an outbreak of local A/H1N1 secondary cases caused by the imported case. The main mode of transmission was personal contact in a short distance without protection, through air and droplet. The locus with poor air ventilation was high risk place. Contact persons should be observed seven days and tested continuously. Infectivity and pathogenicity of the A/H1N1 virus were limited and appeared weakened by generations. Patient's condition was related with persistence and frequency of contact with the infection sources. Enhancing management of contact persons, health education, early diagnose, early treatment and early insulation were effective measures of controling and prenventing the spread A/H1N1.