Combined application of extended depth of field imaging, image stitching and polarized microscopy techniques in identification of .

Objective: As traditional techniques for microscopic identification of Chinese medicines currently lack objective and high-quality reference images, here we developed a systemic procedure to be used in microscopic identification of Chinese medicines, which would lead to more objective, effective and accurate identification process.

Methods: (Jixueteng in Chinese) was used as the specimen in the development of such procedure. Jixueteng samples were microscopically examined in bright- and dark-field microscopy.

Comparative anti-inflammatory effect of curcumin at air-liquid interface and submerged conditions using lipopolysaccharide stimulated human lung epithelial A549 cells.

Background: Inhalation of aerosolized drugs is a promising entry route for rapid and non-invasive therapeutics delivery to the lung. Curcumin exhibits potent anti-inflammatory properties, which are effective for use in lung diseases. The anti-inflammatory properties of curcumin have been widely studied in vitro with cells cultured in submerged conditions, however, the effectiveness using air-liquid interface (ALI) exposure is currently unknown.

A new strategy of studying protein-protein interactions: Integrated strong anion exchange/reversed-phase chromatography/immunoprecipitation coupled with mass spectrometry for large-scale identification of proteins interact with immunoglobulin G in HeLa cells.

Recently, significant research efforts have been devoted to the development of technology for large-scale analysis of protein-protein interactions. Herein, a comprehensive method by coupling the first-dimension strong anion exchange chromatography with the second-dimension reversed-phase liquid chromatography via immunoprecipitation technique and high-resolution mass spectrometry analysis was developed for analyzing protein-protein interactions. After two-dimensional liquid chromatography separation, 108 fractions were obtained in one experiment.

A gold foil covered fused silica capillary tip as a sheathless interface for coupling capillary electrophoresis-mass spectrometry.

A method for the preparation of an on-column ESI emitter used as the sheathless interface for coupling capillary electrophoresis (CE) with mass spectrometry (MS) was developed. The emitter was directly fabricated at the outlet end of the separation capillary which was etched with HF solution to a symmetrical tip. The tip was covered with a small piece of gold foil which was fixed by epoxy resin glue for electrical contact.

Alteration of protein homeostasis mediates the interaction of Pseudomonas aeruginosa with Staphylococcus aureus.

Intracellular protein degradation is essential for the survival of all organisms, but its role in interspecies interaction is unknown. Here, we show that the ClpXP protease of Pseudomonas aeruginosa suppresses its antimicrobial activity against Staphylococcus aureus, a common pathogen co-isolated with P. aeruginosa from polymicrobial human infections.

High performance liquid chromatography-tandem mass spectrometry method for residue determination of 39 plant growth regulators in root and rhizome Chinese herbs.

A simple, rapid, and effective method based on salting-out extraction and LC-MS/MS techniques was developed for the determination of 39 plant growth regulator (PGR) residues in two of the most common root and rhizome Chinese herbs, Codonopsis Radix (Dangshen) and Notoginseng Radix et Rhizoma (Sanqi). The extraction process was performed with acetonitrile-water (5:1) and citrate buffer extraction salt. The performance of the method was validated in accordance with the analytical quality control criteria of SANTE/11813/2017 guidelines.

Isolation and identification of a new sildenafil analogue, hydroxycarbodenafil, found as an adulterant in a health supplement.

During routine screening of illegal adulterants in health supplements, a novel sildenafil analogue was discovered, and subsequently isolated by recrystallization. Its structure was elucidated by extensive analyses of high resolution mass spectrometry (HRMS), one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) data. The analogue was finally determined as hydroxycarbodenafil, featuring a hydroxyethyl group instead of an ethyl group on piperazine ring in comparison with carbodenafil.

Metabolite Profiling and Distribution of Militarine in Rats Using UPLC-Q-TOF-MS/MS.

Militarine, a natural glucosyloxybenzyl 2-isobutylmalate, isolated from , was reported with a prominent neuroprotective effect recently. The limited information on the metabolism of militarine impedes comprehension of its biological actions and pharmacology. This study aimed to investigate the metabolite profile and the distribution of militarine in vivo, which help to clarify the action mechanism further.

The Application of HL60-IL-6 Assay for Pyrogen Detection of CAR-T Cells Products.

Objective: To detect the pyrogen in CAR-T cells product employing the HL60-IL-6 assay.

Methods: The HL60 cells were incubated with CAR-T cells injection or endotoxin standard for 48 hours. After then, the secreted cytokine interleukin-6 (IL-6) from HL60 cells was determined by ELISA.

RNA-seq analysis of testes from flurochloridone-treated rats.

Flurochloridone (FLC) is a widely used herbicide in developing countries. Although the testes are a target organ for FLC in rats, the adverse effects of FLC on testes have not been fully elucidated. To clarify them, we performed RNA-seq analysis using the testes of FLC-treated rats from our previous subchronic toxicity tests.

An integrated approach for global profiling of multi-type constituents: Comprehensive chemical characterization of Lonicerae Japonicae Flos as a case study.

To fully capture the chemo-diversity of medicinal plants is very essential for understanding of their pharmacological activities and guiding scientific quality control. Aiming to facilitate chemical characterization and novel natural products discovery, the present study proposed an integrated approach based on two-dimensional liquid chromatography coupled with quadrupole-Orbitrap mass spectrometer (2D LC/Q-Orbitrap MS). An offline comprehensive two-dimensional (2D) LC system was constructed to cover and separate multi-type constituents by combining hydrophilic interaction chromatography (HILIC) and conventional reversed phase C18.

A novel short-term high-lactose culture approach combined with a matrix-assisted laser desorption ionization-time of flight mass spectrometry assay for differentiating Escherichia coli and Shigella species using artificial neural networks.

Background: Escherichia coli is currently unable to be reliably differentiated from Shigella species by routine matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. In the present study, a reliable and rapid identification method was established for Escherichia coli and Shigella species based on a short-term high-lactose culture using MALDI-TOF MS and artificial neural networks (ANN).

Materials And Methods: The Escherichia coli and Shigella species colonies, treated with (Condition 1)/without (Condition 2) a short-term culture with an in-house developed high-lactose fluid medium, were prepared for MALDI-TOF MS assays.

Qualitative screening and quantitative determination of 569 pesticide residues in honeysuckle using ultrahigh-performance liquid chromatography coupled to quadrupole-Orbitrap high resolution mass spectrometry.

A method for both qualitative screening and quantitative determination of 569 pesticides in honeysuckle was developed based on ultrahigh-performance liquid chromatography (UHPLC) coupled to quadrupole-Orbitrap high resolution mass spectrometry and an in-house executable compound database involving the theoretical masses of precursor and fragment ions and retention times. Different workflows were evaluated, validated and compared including Full MS-SIM, Full MS/ddMS, Full MS/AIF and Full MS/DIA. For qualitative analysis, Full MS/DIA showed relatively low screening detection limits (SDLs) for most pesticides, but its MS identification seemed to be not reliable enough.

Low dose of flurochloridone affected reproductive system of male rats but not fertility and early embryonic development.

Background: Fluorochloridone (FLC) is a widely used herbicide, and its target organs are testes and epididymides. The Globally Harmonized System of Classification and Labelling of Chemicals classified FLC as Level 2-possibly cause fertility or fetal damage (no relevant data support). The maximum residue levels of FLC in processed crops have been reviewed in the latest European Food Safety Authority (EFSA) report in 2018.

Data on the sub-chronic toxicity in rats after 30 days of oral realgar administration and the accumulation and distribution of arsenic species.

These data are related to the research "The accumulation and distribution of arsenic species and association with arsenic toxicity in rats after 30 days of oral realgar administration" (Yi et al., 2019) [1]. These data include the rat body weights, haematology, electrolytes, coagulation and biochemical parameters, and relative organ weights after 30 days of oral administration of realgar, which was consistent with the current OECD guideline "Repeated Dose 28-Day oral Toxicity Study in Rodents".

A composite consisting of sulfo-functionalized magnetic graphene and mesoporous silica for extraction of metformin and glimepiride prior to their determination by liquid chromatography tandem mass spectrometry.

A new sorbent was synthesized for restricted-access matrix solid phase extraction (RAM-SPE) of the diabetes drugs metformin (MET) and glimepiride (Glim). Mesoporous silica layers were placed on FeO-magnetized graphene modified with sulfo-functionalized pore walls (denoted as magG@mSiO-SOH composites). The composites have a large specific surface (173 m·g), appropriate pore sizes (typically 3.

Characterization of unknown impurities in Coenzyme Q using LC-MS and NMR.

During studies on related substances in coenzyme Q (CoQ) active pharmaceutical ingredient (API) and capsules, two impurities (Impurity 1 and Impurity 2) with each content more than 0.1% attracted our attention. Accelerated testing (40 °C /75%RH) and forced degradation of CoQ API and drug products in different stress conditions (base and heat) gave rise to the two unknown impurities.

Rapid and visual detection of berberine hydrochloride based on a water-soluble pyrene derivative.

In this study, a rapid method for the detection of berberine hydrochloride (BRH) was developed based on a water-soluble pyrenyl probe, 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS). This method features low cost, good selectivity, high sensitivity and a fast response. The sensing mechanism of this probe is attributed to the formation of a complex between HPTS and BRH induced by electrostatic interaction and π-π stacking.

[Study on proteins in Guangdilong by nano LC/orbitrap fusion lumos HR-MS].

In the present study,fresh Guangdilong( GD),originating from Pheretima aspergillum,was taken as the object. The total proteins from GD were firstly separated by SDS-PAGE according to their molecular weights and in-gel digestion was then performed.After that,the peptides were analyzed by nano LC/orbitrap fusion lumos high resolution mass spectrometry( nano LC/orbitrap fusion lumos HR-MS).

Multiorgan toxicity induced by EtOH extract of Fructus Psoraleae in Wistar rats.

Background: The fruits of Psoralea corylifolia L. (Fructus Psoraleae, FP) has a long history and a wide range of applications in the treatment of osteoporosis and leukoderma. Although it is well known that FP could cause hepatotoxicity and reproductive toxicity, less is known about its potential toxicity on multiple organs.

Determination of d-limonene in mice plasma and tissues by a new GC-MS/MS method: Comparison of the pharmacokinetics and tissue distribution by oral and inhalation administration in mice.

The aim of the present study was to develop a method based on gas chromatography-tandem mass spectrometry (GC-MS/MS) to determine and quantify the d-limonene in mouse plasma and tissue samples. This new method was validated for the quantification of d-limonene with the linearity ranges 1.0-1000.

The Oral NOAEL of Flurochloridone in Male Wistar Rats in Ninety-Day Subchronic Toxicity Test Was 3mg/kg/day.

A ninety-day toxicity and toxicokinetics of flurochloridone (FLC) were studied in male Wistar rats with oral administration at doses of 3 mg/kg and 10 mg/kg respectively, following the previous study. Apparent toxicity to reproductive system of male rats was still observed at the dose of 10 mg/kg, trace amounts of FLC were still detected 24 hours after administration, testicular weight, epididymal weight and serum testosterone were significantly reduced and sperm abnormalities in epididymis were significantly increased. No abnormalities were found in 3 mg/kg group, it indicated that no-observed-adverse-effect level (NOAEL) of FLC in male rats was 3 mg/kg/day, far below the dose of 20 mg/kg/day reported by European Food Safety Authority (EFSA).

A Comprehensive Multilocus Sequence Typing Scheme for Identification and Genotyping of Strains.

Increasing clinical significance of coagulase-negative staphylococci requires effective methods for species identification and genotyping. In this study, six housekeeping genes (, , , , , and ) with extensive allelic polymorphisms were identified and evaluated to develop a comprehensive multilocus sequence typing (MLST) scheme. Selected primers were capable of amplification of the six loci from all of the 180 strains belonging to 18 different species.

Enzyme immobilized on polyamidoamine-coated magnetic microspheres for α-glucosidase inhibitors screening from Radix Paeoniae Rubra extracts accompanied with molecular modeling.

In this study, a method for direct screening and identification of α-glucosidase inhibitors (AGIs) from extracts of natural products was established based on polyamidoamine (PAMAM) coated magnetic microspheres. A facile route to synthesize the magnetic PAMAM was employed and α-glucosidase was successfully covalently attached to its surface through cross linking of glutaraldehyde. Using the enzyme-loaded magnetic microspheres, potential inhibitors were fished out from crude extracts directly, followed by structure confirmation.