Combination of intramuscular transplantation of autologous mononuclear bone marrow cells with sympathectomy (L2, 3) in patients with peripheral arterial disease(PAD).

BACKGROUND: In most patients with severe, chronic extremity ischemic diseases, intervention or surgical treatment is often not suitable. Combination of intramuscular transplantation of autologous monocular bone marrow cells (AMBMCs) and sympathectomy (L2, 3) has been proved therapeutically beneficial. METHODS: We studied 170 patients (combined group 80, control group 90) with extremity ischemia (TAO, ASO FontaineⅡ,Ⅲ, Ⅳ) between January 2013 and September 2019. RESULTS: In contrast to pre-operation, the walking distance of patients increased significantly (from 61.34 ± 52.23 m to 156.0 ± 32.4 m, p < 0.01), and the ankle-brachial index (ABI) remarkably improved (from 0.28 ± 0.13 to 0.59 ± 0.23, p < 0.05). CONCLUSION: Combined therapy is feasible and effective for patients with peripheral arterial disease (PAD).

Screening of Phosphodiesterase-5 Inhibitors and Their Analogs in Dietary Supplements by Liquid Chromatography-Hybrid Ion Trap-Time of Flight Mass Spectrometry.

An accurate and reliable method based on ion trap-time of flight mass spectrometry (IT-TOF MS) was developed for screening phosphodiesterase-5 inhibitors, including sildenafil, vardenafil, and tadalafil, and their analogs in dietary supplements. Various parameters affecting liquid chromatographic separation and IT-TOF detection were investigated, and the optimal conditions were determined. The separation was achieved on a reversed-phase column under gradient elution using acetonitrile and water containing 0.2% acetic acid at a flow rate of 0.2 mL/min. The chromatographic eluents were directly ionized in the IT-TOF system equipped with an electrospray ion source operating in the positive ion mode. The proposed screening method was validated by assessing its linearity, precision, and accuracy. Sequential tandem MS was conducted to obtain structural information of the references, and the fragmentation mechanism of each reference was proposed for providing spectral insight for newly synthesized analogs. Structural information, including accurate masses of both parent and fragment ions, was incorporated into the MSn spectral library. The developed method was successfully applied for screening adulterated dietary supplement samples.

Discovery of a solvent effect preventing quantitative profiling by matrix-assisted laser desorption/ionization and its treatment.

RATIONALE: In analyte profiling by matrix-assisted laser desorption/ionization (MALDI), drawing a quantitative profile map is an outstanding problem. Recently, we developed a method to quantify an analyte by MALDI, which is needed to solve the problem. Another requirement for quantitative profiling is the quantitative sample-to-matrix analyte transfer, which is investigated in this work. METHODS: MALDI-time-of-flight (TOF) spectra were acquired for samples produced by two methods. In one, a sample solution containing a matrix and an analyte was loaded with a pipet and dried. In the other, a sample was prepared by a consecutive process, i.e., loading-drying of an analyte solution followed by that of a matrix solution. Two different micro-spotters were used in the second method. Various mixtures of organic solvents with water were used to prepare matrix solutions. RESULTS: The organic solvent, matrix, and analyte used in the study did not affect the analyte transfer efficiency, whereas it improved as the water content in the solvent increased. It also improved as the liquid droplet emitted by a micro-spotter got larger. Use of a more polar solvent or a larger droplet increases the contact time between a solution droplet and the sample surface, which seems to be responsible for the improvement in the transfer efficiency. CONCLUSIONS: Sample-to-matrix analyte transfer occurred efficiently when polar solvents and/or large liquid droplets were used to produce solid samples for MALDI profiling with a micro-spotter. A long contact time between the sample surface and a matrix solution droplet is one of the requirements for quantitative profiling. Copyright © 2015 John Wiley & Sons, Ltd.

Performance evaluation of a DNA chip assay in the identification of major genitourinary pathogens.

BACKGROUND: To prevent the recurrence of genitourinary infections and to reduce the risks of their complications, accurate and rapid diagnosis are required. STDetect® Chip is a DNA chip which allows for the simultaneous detection of 13 major genitourinary pathogens in a single vaginal swab or urine specimen. We evaluated the analytical performance of the STDetect® Chip for detecting target pathogens that commonly cause genitourinary infections. METHODS: The target pathogens of the STDetect® Chip are Chlamydia trachomatis, Candida albicans, Enterococcus faecalis, Gardnerella vaginalis, Mycoplasma hominis, Neisseria gonorrhoeae, Mycoplasma genitalium, Ureaplasma urealyticum, Staphylococcus aureus, Klebsiella pneumoniae, Trichomonas vaginalis, and Herpes simplex virus types 1 and 2. Performance of the STDetect® Chip for the detection of target pathogens was evaluated comparing with the result of direct sequencing and conventional multiplex PCR assay. And precision tests for STDetect® Chip were performed with quality control materials. RESULTS: The STDetect® Chip showed high sensitivities (95.1%-100%), specificities (93.4% to 100%), concordance rates (95.0%-100%), positive predictive values (69.8%-100%), and negative predictive values (93.1%-100%) in its identification of 13 target pathogens. The STDetect® Chip had a particularly excellent concordance rate (96.5%) for the 4 major pathogens, C. albicans, G. vaginalis, M. hominis, and U. urealyticum, compared with direct sequencing. Comparing to multiplex PCR assay, STDetect® Chip showed better sensitivity for detecting M. hominis (97.0% vs. 54.5%) and U. urealyticum (93.2% vs. 65.9%). In precision tests, coefficients of variations for signal intensities were ranged from 11.2% to 26.2%. CONCLUSION: The STDetect® Chip showed excellent analytical performance, and its result was in good agreement with that obtained by direct sequencing.

Forsythia fructus inhibits the mast-cell-mediated allergic inflammatory reactions.

Mast cells are key as effector cells in the early phase allergic inflammation and in diverse immunological and pathological processes. Forsythia fructus (F. fructus) has used as a traditional medicine for inflammatory diseases. In the present study, we determined the effect of F. fructus extracts on compound 48/80-induced paw oedema and vascular permeability in vivo. In addition, we investigated in vitro whether F. fructus has inhibitory effects on compound 48/80-induced histamine releases from rat peritoneal mast cells (RPMC), and on phorbol 12-myristate 13-acetate (PMA) plus A23187-induced tumor necrosis factor-alpha (TNF-alpha) releases from human mast cells (HMC-1). In mice orally administrered F. fructus (100 microg/g) for 1 h, compound-48/80-induced oedema and vascular permeability were significantly reduced rather than those receiving intravenous injection of ketotifen, mast cell stabilizer. F. fructus dose-dependently inhibited the histamine release induced by compound 48/80 from RPMCs. Moreover, F. fructus had no cytotoxic effects on cell viability and had inhibitory effects on TNF-alpha secretion from HMC-1. These results suggest that F. fructus is a potential herb medicine for treatment of inflammatory diseases through downmodulating mast cell activation.