An ultrahigh-performance liquid chromatography-tandem mass spectrometry method for quantification of methotrexate and 7-hydroxy-methotrexate and application for therapeutic drug monitoring in patients with central nervous system lymphoma.

A method using ultrahigh-performance liquid chromatography-tandem mass spectrometry was developed, validated, and applied to simultaneously determine plasma methotrexate (MTX) and 7-hydroxy-methotrexate (7-OH-MTX) in 117 patients with central nervous system (CNS) lymphoma. The ion transitions utilized were m/z 455.2 > 308.2 for MTX and m/z 471.2 > 324.1 for 7-OH-MTX. Samples were prepared through protein precipitation using methanol. Chromatographic separation was achieved within 3.0 min on a CMS9030 column (Ruixi, 2.1 × 50 mm, 3 µm) through a gradient elution of methanol and a 10% ammonium acetate solution at a flow rate of 0.4 mL/min. The method demonstrated linearity in the concentration range of 0.05-10 µM for MTX and 0.25-50 µM for 7-OH-MTX. The intra- and inter-day inaccuracy ranged from -7.38% to 7.83%, and the imprecision was less than 6.00% for both analytes. The recovery and matrix effect normalized by the internal standard (MTX-D3 ) remained consistent. Both analytes remained stable under nine different storage conditions. In patients with CNS lymphoma, MTX levels at 12 h and 7-OH-MTX levels at 12, 36, and 60 h after dosing in individuals with impaired renal function were significantly higher compared with those with normal renal function. 7-OH-MTX could potentially serve as a superior indicator for nephrotoxicity compared with MTX.

Key factors influencing on vegetation restoration in the gullies of the Mollisols.

Natural vegetation restoration (NVR) highly relates to the development of gully erosion, and is mainly determined by both the soil properties and species competition in the gullies. However, it is still not clear what are the key factors influencing on the vegetation restoration in the gullies with the poor soil properties (e.g. low soil organic matter and nutrients) under the special hydrological process (e.g. high runoff intensity and long flow duration). In this study, soil total organic carbon (SOC), total nitrogen (TN), available nitrogen (AN), ammonium nitrogen (NH4+-N), nitrate nitrogen (NO3--N), total phosphorus (TP), available phosphorus (AP), pH, soil moisture (SM) were investigated, and both regression and structural equation model analysis were used for detecting how soil properties and species competition influence the herbaceous plants restoration in the poor quality of Mollisols in gullies of Northeast China. The results show that, (1) influence of NH4+-N, AN, TN, pH on biomass was stronger in 0-10 cm than that in 10-20 cm soil depth, opposite was stronger in 10-20 cm than that in 0-10 cm soil depth for NO3--N, SOC and SM (P < 0.05). (2) NH4+-N, NO3--N, AN, TN, SOC, pH, C:N were all negative, while SM was positive to plant biodiversity in soil layers (P < 0.05). (3) SOM mainly mediates the TN and NH4+-N and then directly or indirectly influences on biodiversity and biomass, and P changed the species richness when AP >20 mg kg-1 in 10-20 cm soil depth. (4) Vegetation restoration was mainly determined by the dynamics of Elymus dahuricus Turcz. firstly, and then by Leymus chinensis(Trin.) Tzvel. at the early of vegetation restoration. Generally, the heterogeneity of SOC and SM in soil layers and AP in deep soil layer was the key factors determining NVR in the gullies of Mollisols watershed. At the end of paper, the NVR process in Moillosols in gullies was classified as four stages, and each stage was depicted in detail.

Development and validation of an LC-MS/MS method for the determination of tigecycline in human plasma and cerebrospinal fluid and its application to a pharmacokinetic study.

Tigecycline (TGC) is an important antibiotic in treating various drug-resistant bacteria. The dosage regimen for cerebral intraventricular TGC is still unknown. The aim of the study was to develop and validate liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for the determination of TGC in human plasma and cerebrospinal fluid (CSF) to obtain an applicable regimen. The ion transitions under ESI positive model were performed at m/z 586.3 > 513.2 and m/z 595.3 > 514.3 for TGC and d9-TGC internal standard (IS). For plasma and CSF samples, the calibration curve of TGC was linear within the ranges 25-2000 and 250-100,000 ng/mL; the IS normalized matrix effect was within the ranges 96.46-101.06% and 101.13-103.58%, respectively, for all. TGC was stable under all tested conditions. The patient received 1 mg intraventricular and 49 mg intravenous administration of TGC. The AUC0-12 in plasma and CSF calculated according to our noncompartment model were 4713 and 23,0238 h ng/mL, respectively. Given our findings cerebral intraventricular TGC may be a choice for clinicians to treat drug-resistant Gram-negative bacterial-induced meningitis and the safety and efficacy of this administration route warrants further study.

Determination of pseudo-ginsenoside GQ in human plasma by high performance liquid chromatography-tandem mass spectrometry.

A specific, sensitive and rapid method based on high performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) was developed for the determination of pseudo-ginsenoside GQ in human plasma. Liquid-liquid extraction was used to isolate the analyte from biological matrix followed by injection of the extracts onto a C8 column with isocratic elution. Detection was carried out on a triple quadrupole tandem mass spectrometer (API-4000 system) in multiple reaction monitoring mode using negative electrospray ionization. The mobile phase consisted of methanol-10 mM ammonium acetate (90:10, v/v) and the flow rate was 0.3 mL/min. The method was validated over the concentration range of 5.0-5000.0 ng/mL for plasma. Inter- and intra-day precisions (relative standard deviation) were all within 15% and the accuracy (relative error) was ≤ 9.4%. The lower limit of quantitation was 5.0 ng/mL. The pseudo-ginsenoside GQ was stable after 8 h at room temperature, 24 h at autosampler and three freeze-thaw cycles (from -30 to 25 °C). The method was successfully applied to the pharmacokinetic study of pseudo-ginsenoside GQ in healthy Chinese volunteers.

A UHPLC-MS/MS method for the simultaneous determination of vancomycin, norvancomycin, meropenem, and moxalactam in human plasma and its clinical application.

We developed an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to determine four antibacterial drugs in human plasma for clinical usage. Samples were prepared using protein precipitation with methanol. Chromatographic separation was accomplished in 4.5 min on a BEH C18 column (2.1 × 50 mm, 1.7 µm) using a gradient elution of methanol and water (containing 7.71 g/L concentrated ammonium acetate, adjusted to pH 6.5 with acetic acid) at a flow rate of 0.4 mL/min. Positive electrospray was used for ionization. The method was linear in the concentration range 1-100 µg/mL for vancomycin, norvoncomycin, and meropenem; and 0.5-50 µg/mL for R-isomer of moxalactam and S-isomer of moxalactam. For all analytes, the intra- and inter-day accuracies and precisions were -8.47%-10.13% and less than 12%, respectively. The internal standard normalized recoveries and matrix effect were 62.72%-105.78% and 96.67%-114.20%, respectively. All analytes were stable at six storage conditions, with variations of less than 15.0%. The method was applied in three patients with central nervous system infection. The validated method might be useful for routine therapeutic drug monitoring and pharmacokinetic study.

Trends in research related to vaccine and cancer prevention from 1992 to 2022: A 30-years bibliometric analysis.

Vaccines may play an important role in cancer prevention. This bibliometric study in the field of vaccine and cancer prevention is designed to evaluate key research advances, identify existing deficiencies, and provide reference for future investigations. A total of 2916 original articles published in English from 1992 to 2022 were extracted from the Web of Science core collection. America (1,277) and the National Cancer Institute (82) were the most productive country and institution in this field, respectively. Vaccine was not only the most co-cited journal but also the most influential. Garland SM was the most prolific author, and Bosch FX was the most influential co-cited author. The keywords "cervical cancer" had the highest frequency. "Nanovaccines," "vaccine acceptance" and "coverage" were the new research hotspots in this field. Currently, although an increasing number of publications involve vaccine and cancer prevention studies, most of them are related to cervical cancer, and few other cancers, suggesting the need to investigate other cancer prevention vaccines. The promising research hotspots, such as nanovaccines, vaccine acceptance, and vaccine coverage should be the focus of investigation. The study provides the current status and trends in clinical research on vaccine and cancer prevention, enabling researchers to identify hotspots and explore new study directions. In the future, vaccines are expected to play a key role in multiple cancer prevention.

What is the context? Currently, cancer is the leading cause of death worldwide and poses a significant burden due to increasing incidence, high mortality, and heavy economic costs. One effective way to prevent cancers is by using cancer prevention vaccines such as the human papillomavirus (HPV) vaccine. By preventing an infection that can progress to cancer and reducing the morbidity and economic burden, HPV vaccines have played a positive role in the fight against cervical cancer. The potential of vaccines as a cancer prevention strategy is significant. However, the shortcomings, progress, hotspots and frontiers of global research in the field of vaccine and cancer prevention remain unclear.What is new? In this study, a bibliometric analysis was conducted from multiple aspects such as countries, institutions, authors, co-cited authors/journals/references and keywords. The analysis was designed to evaluate the important research progress in vaccine and cancer prevention, to explore the research hotspots and frontiers, and to identify existing problems and deficiencies. It provides a valuable reference for future research directions.What is the impact? Excluding cervical and liver cancer, few other cancers have been investigated in the field of vaccine and cancer prevention, suggesting the need for intensive studies focusing on other cancer prevention vaccines. Key areas for investigation include nanovaccines, vaccine acceptance, and vaccine coverage. In the future, vaccines are expected to play a more important role in prevention of cancers beyond cervical cancer prevention.

Causal relationship between spondylarthritis and stroke in a European population: a two sample Mendelian randomization study.

Background: Observational studies have found an increased risk of stroke in patients with spondyloarthritis, but the results are susceptible to reverse causality and confounders. Therefore, the study aimed to further explore the association between spondyloarthritis and different subtypes of stroke by using a two sample Mendelian randomization (MR) analysis. Methods: Genetic instrumental variables for spondyloarthritis were identified using summary level data from a genome-wide association study involving 201,581 people. Summary statistics from the Multiancestry Genome-wide Association Study of Stroke Consortium were used to obtain genetic data on stroke. There was no sample overlap between the exposure and outcome datasets. Inverse-variance weighted was considered the primary MR method for causal analysis. Heterogeneity, pleiotropy and sensitivity analyses were performed to ensure robustness, and single nucleotide polymorphism (SNP) with potential confounders was further screened in the PhenoScanner database to better evaluate the stability of our study. Results: One SNP (rs1065045) was excluded due to schizophrenia. After excluding SNP (rs1065045), results of the second MR analysis were slightly different from the first, which were considered as the final result: a significant positive causality between spondyloarthritis and cardioembolic stroke (OR=1.296, 95% CI:1.094-1.534, p=0.003); a possible positive causality between spondyloarthritis and any stroke (OR=1.082, 95% CI:1.016-1.152, p=0.013)/any ischemic stroke (OR=1.086, 95% CI:1.013-1.163, p=0.020); no significant/possible causality between spondyloarthritis and small vessel stroke (OR=1.168, 95% CI:0.993-1.375, p=0.061). Insufficient power may be one possible reason why a causality was not observed between spondyloarthritis in our study. Conclusions: This study suggests that the possible causative effects of spondyloarthritis predicted by genetics on stroke may be limited to any stroke, any ischemic stroke, and cardioembolic stroke, especially the last.

Utilization of glucagon-like peptide-1 receptor agonists in children and adolescents in China: a real-world study.

Background: Glucagon-like peptide-1 receptor agonists (GLP-1RAs) have been widely used in treating type 2 diabetes mellitus (T2DM) and obesity in adults, but scientific research about the indication in children and adolescents is scarce. The current study aims to explore the prescriptions of GLP-1RAs in children and adolescents in China and to evaluate its rationality. Methods: GLP-1RA prescriptions of children and adolescents were retrospectively obtained from the Hospital Prescription Analysis Cooperative Project. The study extracted information on patient's demographic characteristics, monotherapy and combination therapy of GLP-1RAs, and trends in GLP-1RA usage from 2016 to 2021. The rationality of GLP-1RA prescriptions was comprehensively assessed based on the indications approved by China National Medical Products Administration (NMPA), the U.S. Food and Drug Administration (FDA), European Medicines Agency (EMA), Pharmaceuticals and Medical Devices Agency (PMDA), and published randomized controlled trials (RCTs). Results: A total of 234 prescriptions from 46 hospitals were included, with a median age of 17 years old. The majority of patients were diagnosed with overweight/obesity or prediabetes/diabetes, accounting for 43.59% and 46.15%, respectively. There were 88 patients on GLP-1RA monotherapy. GLP-1RAs plus metformin was the most common combination therapy (38.89%). 12.39% of patients were found a co-administration with orlistat. The share of overweight/obesity prescriptions increased from 27% in 2016 to 54% in 2021, whereas prediabetes/diabetes prescriptions declined from 55% to 42%. The prescriptions were divided into appropriate and questionable groups according to the diagnosis, and the potentially questionable prescription was related to age (p = 0.017), department visited (p = 0.002), and any hospitalization (p < 0.001). Conclusions: This study described the prescribing of GLP-1RAs in children and adolescents. Our findings indicated that the utilization of GLP-1RAs has increased from 2016 to 2021. There was a strong basis for administering GLP-1RAs in overweight/obesity and prediabetes/diabetes, whereas the evidence was insufficient in other conditions. It is crucial to demand robust and sustained efforts to enhance the awareness of the safety of utilization of GLP-1RAs in children and adolescents.

Real-world national trends and socio-economic factors preference of sodium-glucose cotransporter-2 inhibitors and glucagon-like peptide-1 receptor agonists in China.

Backgrounds: Robust evidence have demonstrated the beneficial effect of Sodium-glucose cotransporter-2 inhibitors (SGLT2i) and glucagon-like peptide-1 receptor agonists (GLP-1RA) in T2D patients with cardiovascular diseases and chronic kidney disease. Multiple studies analyzed patterns and predictors of SGLT2i and GLP-1RA in the US, Europe and worldwide. However, there is no study about the utilization of these two classes of drugs in real-world in China. Method: A total of 181743 prescriptions of SGLT2i and 59720 GLP-1RA were retrospectively pooled from Hospital Prescription Analysis Cooperation Project from 2018 to 2021. The social-economic characteristics of patients and prescribers, including age, gender, residency, hospital level, insurance type, department visited, and payment amount, were collected and analyzed to study trends and risk factors associated with preference among two antidiabetics. Results: Annual number of prescriptions of SGLT2i significantly increased to approximately 140 folds, while GLP-1RA increased to about 6.5 folds. After adjustment for socio-economic information, several patients or physician characteristics were positively associated with the preference of GLP-1RA, including female gender (OR 1.581, 95% CI 1.528-1.635), residents in second-tier cities (OR 1.194, 95% CI 1.148-1.142), visiting primary or secondary hospital level (OR 2.387, 95% CI 2.268-2.512); while other factors were associated with the preference of SGLT2i, including older adults (OR 0.713, 95% CI 0.688-0.739), uncovered by insurance (OR 0.310, 95% CI 0.293-0.329), visiting other departments compared with endocrinology. In addition, the share of SGLT2i and GLP-1RA was low but in an increasing tendency. Conclusions: SGLT2i and GLP-1RA prescription significantly increased from 2018 to 2021. The socio-economic risk factors in choosing SGLT2i or GLP-1RA highlight an effort required to reduce disparities and improve health outcomes.

CTLA-4 Facilitates DNA Damage-Induced Apoptosis by Interacting With PP2A.

Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) plays a pivotal role in regulating immune responses. It accumulates in intracellular compartments, translocates to the cell surface, and is rapidly internalized. However, the cytoplasmic function of CTLA-4 remains largely unknown. Here, we describe the role of CTLA-4 as an immunomodulator in the DNA damage response to genotoxic stress. Using isogenic models of murine T cells with either sufficient or deficient CTLA-4 expression and performing a variety of assays, including cell apoptosis, cell cycle, comet, western blotting, co-immunoprecipitation, and immunofluorescence staining analyses, we show that CTLA-4 activates ataxia-telangiectasia mutated (ATM) by binding to the ATM inhibitor protein phosphatase 2A into the cytoplasm of T cells following transient treatment with zeocin, exacerbating the DNA damage response and inducing apoptosis. These findings provide new insights into how T cells maintain their immune function under high-stress conditions, which is clinically important for patients with tumors undergoing immunotherapy combined with chemoradiotherapy.

Cryptogenic organizing pneumonia masquerading as lung carcinoma: A case report and review of the literature.

Cryptogenic organizing pneumonia (COP) is a rare pulmonary disorder of unknown etiology. COP with hemoptysis as the primary presenting symptom has rarely been reported. The present study reported a case of COP that resembled lung carcinoma with hemoptysis as the only clinical symptom. The patient recovered well following thoracoscope surgery. A literature review of 119 COP cases between 1995 and 2015 was presented. Cough, fever and dyspnea were the most common clinical manifestations. The most common imaging manifestations were multiple or single consolidation, lung nodules, migratory sign, reversed halo sign, and multiple ground-glass opacity. A total of 3 cases exhibited COP accompanied by lung cancer. Glucocorticoids were effective for the majority of cases and invasive surgeries were implemented in most cases. The majority of cases recovered or relieved, and the prognosis of COP was relatively good. COP was easily confused with lung tumor and it is necessary to make differential diagnosis between COP and lung cancer. Invasive surgery should be avoided when possible to avoid or reduce patient trauma.

Identification of two immortalized cell lines, ECV304 and bEnd3, for in vitro permeability studies of blood-brain barrier.

To identify suitable cell lines for a mimetic system of in vivo blood-brain barrier (BBB) for drug permeability assessment, we characterized two immortalized cell lines, ECV304 and bEnd3 in the respect of the tightness, tight junction proteins, P-glycoprotein (P-gp) function and discriminative brain penetration. The ECV304 monoculture achieved higher transendothelial electrical resistance (TEER) and lower permeability to Lucifer yellow than bEnd3. However, co-culture with rat glioma C6 cells impaired the integrity of ECV304 and bEnd3 cell layers perhaps due to the heterogeneity among C6 cells in inducing BBB characteristics. The immunostaining of ZO-1 delivered distinct bands along cell borders on both cell lines while those of occludin and claudin-5 were diffused and weak. P-gp functionality was only proved in bEnd3 by Rhodamine 123 (R123) uptake assay. A permeability test of reference compounds displayed a similar rank order (digoxin < R123 < quinidine, verapamil < propranolol) in ECV304 and bEnd3 cells. In comparison with bEnd3, ECV304 developed tighter barrier for the passage of reference compounds and higher discrimination between transcellular and paracellular transport. However, the monoculture models of ECV304 and bEnd3 fail to achieve the sufficient tightness of in vitro BBB permeability models with high TEER and evident immunostaining of tight junction proteins. Further strategies to enhance the paracellular tightness of both cell lines to mimic in vivo BBB tight barrier deserve to be conducted.

UPLC-MS/MS Analysis of Methotrexate in Human Plasma and Comparison with the Fluorescence Polarization Immunoassay.

Methotrexate (MTX) plasma concentration is routinely monitored to guide the dosage regimen of rescue drugs. This study aims to develop and validate an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for plasma MTX analysis, and to establish its agreement with the fluorescence polarization immunoassay (FPIA) in patients with high-dose MTX therapy. Separation was achieved by gradient elution with methanol and water (0.05% formic acid) at 40°C with a run time of 3 min. The intra- and inter-day inaccuracy and imprecision of the UPLC-MS/MS method were -4.25 to 3.1 and less than 7.63%, respectively. The IS-normalized recovery and matrix effect were 87.05 to 92.81 and 124.43 to 134.57%. The correlation coefficients between UPLC-MS/MS and FPIA were greater than 0.98. The UPLC-MS/MS method was in agreement with the FPIA at high levels of MTX (1.0 - 100 µmol/L), but not at low levels (0.01 - 1.0 µmol/L). Further studies are warranted to confirm these results.