[Determination of 145 pharmaceuticals and personal care products in eleven categories in water by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry].

Pharmaceuticals and personal care products (PPCPs) are emerging contaminants frequently detected in aquatic environments at trace levels. These chemicals have diverse structures and physicochemical properties and includes pharmaceuticals like antibiotics, antihypertensive drugs, antiviral drugs, and psychotropic drugs that are widely used in large quantities worldwide. Considering the large number of pharmaceuticals currently in usage, it is crucial to establish a priority list of PPCPs that should be monitored and/or treated first. An accurate understanding of the occurrence and levels of PPCPs in aquatic environments is essential for providing objective materials for monitoring these emerging contaminants. Therefore, accurate, efficient, sensitive, and high-throughput screening techniques need to be established for determining and quantifying PPCPs. This study developed a method for the determination of 145 PPCPs (grouped into eleven categories: antibiotics, antihypertensive drugs, antidiabetic drugs, antiviral drugs, ß-receptor agonists, nitroimidazoles, H2 receptor antagonists, psychotropic drugs, hypolipidemic drugs, non-steroidal anti-inflammatory drugs, and others) in water. The method was based on large volume direct injection without sample enrichment and cleanup and used ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-MS/MS). Water samples were collected and filtered through a 0.22-µm regenerated cellulose (RC) filter membrane. Subsequently, Na2EDTA was added to the samples to adjust their pH to 6.0-8.0. Internal standards were mixed with the solutions, and because of the addition of Na2EDTA, the interference of metal ions could be eliminated in the determination of compounds, especially for tetracycline and quinolone antibiotics. Among the six filter membranes tested in this study (PES, PFTE-Q, PFTE, MCE, GHP, and RC), RC filter membranes were screened for water sample filtration. The UHPLC-MS/MS parameters were optimized by comparing the results of various mobile phases, as well as by establishing the best instrumental conditions. The 145 PPCPs were separated using an Phenomenex Kinetex C18 column (50 mm×3 mm, 2.6 µm) via gradient elution. The mobile phases were 0.1% (v/v) formic acid aqueous solution containing 5 mmol/L ammonium formate and acetonitrile for positive ion modes, 5 mmol/L aqueous solutions of ammonium formate and acetonitrile for negative ion modes. The samples were quantified using the scheduled multiple reaction monitoring (scheduled-MRM) mode with electrospray ionization in positive and negative ion modes. A standard internal calibration procedure was used to calculate contents of sample. The established method was systematically verified, and it demonstrated a good linear relationship. The average recoveries of the 145 PPCPs at the three spiked levels were in the range of 80.4%-128% with relative standard deviations (RSDs, n=6) of 0.6%-15.6%. The method detection limits (MDLs) ranged from 0.015 to 5.515 ng/L. Finally, the optimization method was applied to analyze the 145 PPCPs in 11 surface water samples and 6 drinking water samples. Overall, 93 (64%) out of the 145 analytes were detected. The total contents of the PPCPs in surface water samples ranged from 276.9 to 2705.7 ng/L. The detection frequencies of antidiabetic, antiviral, and psychotropic drugs were 100%. The total contents of the PPCPs in drinking water samples ranged from 140.5 to 211.5 ng/L, and antibiotics, antidiabetic drugs, and antiviral drugs comprised the largest proportion of analytes (by mass concentration) in drinking water samples. Our method exhibited high analytical speed and high sensitivity. It is thus suitable for the trace analysis and determination of the 145 PPCPs in environmental water and showed improved detection efficiency for PPCPs in water, indicating that it has a high potential for practical applications. This study can extend technical support for further pollution-level analysis of PPCPs in water and provide an objective basis for environmental management.

SWATH-F: A Novel Nontarget Strategy Based on the SWATH-MS Deconvolution Method Assisting in Annotating PFAS Homologues in Multisample Studies.

In order to identify emerging per- and polyfluoroalkyl substances (PFASs) and their alternatives in the environment or population, we need to perform extensive profiling of PFASs to determine their distribution in samples. The sequential window acquisition of all theoretical fragment-ion spectra (SWATH mode) is capable of obtaining a wide range of MS2 spectra but is difficult for direct identification of PFASs due to its complex MS2 spectra, and the nontarget screening method is difficult to identify due to its lack of a priori information. In this study, we demonstrated the great potential of SWATH-F, a nontarget fragment-based homologue screening method in combination with the SWATH-MS deconvolution, for detecting PFASs. We evaluated the application of SWATH-F to gradient spiked samples and real population serum samples, compared it with nontarget homologue screening in the information-dependent acquisition mode (IDA mode), and obtained better results for SWATH-F with 276% improvement (IDA:17 PFASs, SWATH-F: 64 PFASs) in identification. In addition, we automated the screening and identification process of SWATH-F to facilitate its use by researchers. SWATH-F is freely available on GitHub (https://github.com/njuIrene/SWATH-F) under an MIT license.

MicroRNA-411-3p motivates methotrexate's cellular uptake and cytotoxicity via targeting Yin-yang 1 in leukemia cells.

This study aimed to figure out how microRNA (miR)-411-3p's impacts on methotrexate (MTX)'s cellular uptake and cytotoxicity in acute lymphoblastic leukaemia (ALL) CEM-C1 cells by targeting Yin-yang 1 (YY1). miR-411-3p and YY1 were detected by RT-qPCR or Western blot. Intracellular MTX concentration was measured by enzyme-linked immunosorbent assay. Cell viability and apoptosis were evaluated by CCK-8, clonal formation assay, and flow cytometry. Verification of miR-411-3p and YY1's targeting link was manifested. It came out that miR-411-3p mimic or si-YY1 elevated intracellular MTX, MTX-induced cytotoxicity and apoptosis rate in CEM-C1. However, the inverse results were noticed in cells introduced with miR-411-3p inhibitor or oe-YY1. Meanwhile, it was found that cell relative luciferase activity was reduced after co-transfection of miR-411-3p mimic with YY1-WT, indicating that miR-411-3p targeted YY1. Elevation of YY1 could turn around elevating miR-411-3p's impacts on MTX's cellular uptake and cytotoxicity in CEM-C1 cells. These findings convey that miR-411-3p motivated MTX's cellular uptake and cytotoxic impacts via targeting YY1 in leukemia cells. This study is helpful for learning about the mechanisms underlying MTX responses in ALL patients.

Investigation on the Influencing Factors Related to Quality of Life of Adult Epilepsy Patients in Wenzhou, China, Based on Structural Equation Model.

Objective: The aim of the study is to investigate the influencing factors of quality of life in adult patients with epilepsy in Wenzhou in China. Methods: A total of 190 patients who visited our hospital from July 2019 to February 2021 were included in the study. Demographic data and disease status were collected. Moreover, QOLIE-31, PSQI, ESS, HAMD-17, and GAD-7 were used in the questionnaire survey. Structural equation model fitting was used to analyze the influencing factors of quality of life in adult patients with epilepsy. Results: The scores of the dimension of onset worry in men were greater than those of women (P = 0.049), and the scores of the dimension of life satisfaction were lower than women (P = 0.047). The scores of cognitive function decreased with age (P = 0.047). The scores of quality of life of unemployed and drinking patients significantly decreased (P < 0.05). The score of quality of life positively correlated with good economic status and family relations (P < 0.05). The score of emotional health increased first and then decreased with the course of the disease. With the decrease in seizure frequency and the extension of months without a seizure, the score of quality of life gradually increased. Furthermore, the structural equation model showed that health status was directly correlated to the quality of life of patients with epilepsy. Conclusion: Male, unemployment, drinking, older age, and disease are negatively related to the quality of life in patients with epilepsy. However, good economic status, good family relations, and good colleague relationships are positively related to the quality of life.

[Determination of 42 antibiotic residues in seven categories in water using large volume direct injection by ultra high performance liquid chromatography-triple quadrupole mass spectrometry].

Antibiotics are emerging contaminants that have recently attracted attention. They have been detected in natural water and pose health concerns owing to potential antibiotic resistance. Antibiotics are ubiquitous in aquatic environments, with a wide spectrum and trace levels. It is difficult to detect all types of antibiotics with completely different physicochemical properties. Solid phase extraction (SPE) is a common sample preparation procedure. For a fast and high-throughput continuous on-line analysis of these emerging contaminants, a method for the determination of 42 antibiotics (grouped into seven categories: sulfonamides, fluoroquinolones, lincosamides, macrolides, tetracyclines, cephalosporins, and chloramphenicols) in environmental water was developed based on ultra high performance liquid chromatography combined with tandem mass spectrometry (UHPLC-MS/MS) involving large volume direct injection without sample enrichment and cleanup. The collected water samples were filtered through a 0.22-µm filter membrane, their pH levels were adjusted to 6.0-8.0 after adding Na2EDTA, and then the solutions were mixed with an internal standard. The addition of Na2EDTA contributed to the release of tetracyclines and fluoroquinolones from the metal chelate. Improved recoveries were observed for all the compounds when the pH of the aqueous solution was set at 6.0-8.0. The optimized UHPLC conditions were as follows: chromatographic column, Phenomenex Kinetex C18 column (50 mm×30 mm, 2.6 µm); mobile phase, acetonitrile and 0.1% (v/v) formic acid aqueous solution; flow rate, 0.4 mL/min; injection volume, 100 µL. In the UHPLC-MS/MS experiment, chloramphenicol, thiamphenicol, and florfenicol were analyzed in the negative ionization scheduled multiple reaction monitoring mode (scheduled-MRM), while the other 39 antibiotics were analyzed in the positive scheduled-MRM mode. This acquisition method improved the response of each target compound by dividing the time of the analysis test cycle and scanning the ion channels of chromatographic peaks at different time periods. The ionspray voltage was set at 5500 and -4500 V in positive and negative modes, respectively. The source temperature for both ionization modes was set at 500 ℃, which was optimized to improve the sensitivity. Instrumental parameters like collision energy and declustering potential were also optimized. Good linearity was observed for all the tested antibiotics, with a correlation coefficient (r) greater than 0.995. The method detection limits (MDLs) were 0.015-3.561 ng/L. The average recoveries ranged from 80.1% to 125%, while the relative standard deviations (RSDs) were between 0.8% and 12.2%. The method was successfully applied to the determination of 10 source water samples and 5 tap water samples. Twelve antibiotics, viz. sulfachloropyridazine, sulfadiazine, sulfamethazine, sulfamethoxazole, sulfisomidine, clindamycin, lincomycin, roxithromycin, clarithromycin, erythromycin, thiamphenicol, and forfenicol, were detected in the 10 water samples with a detection frequency of 100%. The total antibiotic content in each sample ranged from not detected to 80.3 ng/L. Lincosamides and chloramphenicols were the predominant antibiotics in the water samples, with contents in the ranges of 3.83-13.7 and 4.23-33.6 ng/L, respectively. Therefore, the large volume direct injection method exhibited good performance in terms of MDL and recovery compared to standard methods and those reported previously. Compared with traditional pretreatment methods, the large volume direct injection method is simpler, more rapid, more precise, and more accurate. It is a viable alternative to SPE, and can be used for the determination of the 42 antibiotics at trace levels in cleaner water bodies, such as surface water, groundwater, and tap water.

Observation of the characteristics of the natural course of Bietti crystalline dystrophy by fundus fluorescein angiography.

BACKGROUND: Bietti crystalline dystrophy (BCD) is an autosomal recessive genetic disorder that causes progressive vision loss. Here, 12 patients were followed up for 1-5 years with fundus fluorescein angiography (FFA) to observe BCD disease progression. METHODS: FFA images were collected for 12 patients with BCD who visited our clinic twice or more over a 5-year period. Peripheral venous blood was collected to identify the pathogenic gene related to the clinical phenotype. RESULTS: We observed two types in FFA images of patients with BCD. Type 1 showed retinal pigment epithelium (RPE) atrophy in the macular area, followed by choriocapillaris atrophy and the subsequent appearance of RPE atrophy appeared at the peripheral retina. Type 2 showed RPE atrophy at the posterior pole and peripheral retina, followed by choriocapillaris atrophy around the macula and along the superior and inferior vascular arcades and the nasal side of the optic disc. The posterior and peripheral lesions of both type 1 and type 2 BCD subsequently extended to the mid-periphery; finally, all the RPEs and choriocapillaris atrophied, exposing the choroid great vessels, but type 2 macular RPE atrophy could last longer. CONCLUSIONS: The characterization of two different types of BCD development provides a better understanding of the phenotype and the progression of the disease for a precise prognosis and prediction of pathogenesis.

Development of a liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for simultaneous determination of epigallocatechin-3-gallate, silibinin, and curcumin in plasma and different tissues after oral dosing of Protandim in rats and its application in pharmacokinetic and tissue distribution studies.

Protandim is an over-the-counter herbal dietary supplement. The key components of Protandim, i.e., epigallocatechin-3-gallate (EGCG), silibinin (SIL), and curcumin (CUR) were simultaneously analyzed through a liquid chromatography-tandem mass spectrometric (LC-MS/MS) method in plasma and different tissues after administration of Protandim in rats. The developed and validated method was employed to assess the pharmacokinetic profiles and the accumulation of EGCG, SIL, CUR in rat plasma and tissue homogenates. The plasma and tissue homogenates were subjected to liquid-liquid extraction and separated on a Hypurity C18 column (50 × 4.6 mm) with a gradient elution of water and acetonitrile. Mass spectrometric detection was performed in the multiple reaction monitoring mode (MRM) following the transitions: m/z 457.3/169.3, m/z 481.3/125.0, m/z 367.3/149.3 and m/z 609.4 /300.2 for EGCG, SIL, CUR, and RU (rutin), respectively. The concentrations of all the analytes in the range from 2 to 1000 ng/mL showed linear relationships with respective peak areas in different matrices. For all matrices, the values of inter-day and intra-day precisions and accuracies were less than 10.3% of the nominal concentration. The matrix effect, extraction recovery, dilution integrity, and stability values were all within acceptable levels. This method was successfully applied for determining the pharmacokinetics and tissue distribution of the components in rats after the intragastrical administration of a single-dose (364.5 mg/kg) or multiple-doses (1458 mg/kg) of Protandim. The data showed that EGCG, SIL, and CUR did not accumulate in rats after multiple doses of Protandim, and the three main components were distributed mainly in the small intestine.

Psychotherapy combined with drug therapy in patients with category III chronic prostatitis/chronic pelvic pain syndrome: A randomized controlled trial.

OBJECTIVES: To explore the efficacy of psychotherapy combined with drug therapy in patients with category III chronic prostatitis/chronic pelvic pain syndrome. METHODS: A total of 156 patients with category III chronic prostatitis/chronic pelvic pain syndrome were randomly divided into two groups: the control group of 78 patients receiving routine medication; and the intervention group of 78 patients receiving psychological intervention therapy combined with routine medications. Treatment courses were for 3 months. The end-points were the response rate of the National Institutes of Health Chronic Prostatitis Symptom Index, International Index of Erectile Function-5, Self-Rating Anxiety Scale, Self-Rating Depression Scale and expressed prostatic secretion-white blood cells. RESULTS: After 3 months, the average scores of the National Institutes of Health Chronic Prostatitis Symptom Index decreased to 10.1 ± 5.0 in the control group compared with 14.1 ± 4.9 in the intervention group; thus, significant differences were observed between the two groups in the study (P < 0.001). The average scores of the International Index of Erectile Function-5 were improved in the two groups, but compared with the control group, a more marked improvement was detected in the psychological intervention group, and there was a significant difference between the two groups (P < 0.001). There was significant difference between the two groups in terms of the Self-Rating Anxiety Scale and Self-Rating Depression Scale scores (P < 0.001). Expressed prostatic secretion-white blood cell counts significantly decreased to 4.4 ± 3.5 in the control group compared with 9.8 ± 3.4 in the intervention group (P < 0.001). CONCLUSIONS: Psychological intervention therapy can effectively improve the psychological status and sexual function in patients with category III chronic prostatitis/chronic pelvic pain syndrome than the routine medication.

Amphiphilic dendritic nanomicelle-mediated co-delivery of 5-fluorouracil and doxorubicin for enhanced therapeutic efficacy.

Combination cancer therapy has attracted considerable attention due to its enhanced antitumor efficacy and reduced toxicity granted by synergistic effects over monotherapy. The application of nanotechnology is expected to achieve coencapsulation of multiple anticancer agents with enhanced therapeutic efficacy. Herein, a unique nanomicelle based on amphiphilic dendrimer (AmD) consisting of a hydrophilic polyamidoamine dendritic shell and a hydrophobic polylactide core is developed for effectively loading and shuttling 5-fluorouracil (5-Fu) and doxorubicin (Dox). The yielded drug-encapsulated dendritic nanomicelle (5-Fu/Dox-DNM) has a modest average size of 68.6 ± 3.3 nm and shows pH-sensitive drug release manner. The parallel activity of 5-Fu and Dox show synergistic anticancer efficacy. The IC50 value of 5-Fu/Dox-DNM toward human breast cancer (MDA-MB-231) cells was 0.25 µg/mL, presenting an 11.2-fold and 6.1-fold increase in cytotoxicity compared to Dox-DNM and 5-Fu-DNM, respectively. Furthermore, 5-Fu/Dox-DNM significantly inhibits the progression of tumor growth in the MDA-MB-231 xenograft tumor mice model. In conclusion, we have demonstrated that our AmD-based combination therapeutic system has promising potential to open an avenue for coencapsulation of multiple chemotherapeutic agents to promote superior anticancer effect.