两种药物膀胱灌注治疗间质性膀胱炎的临床疗效观察

目的 观察并比较透明质酸钠、碱化利多卡因联合肝素膀胱灌注治疗膀胱疼痛综合征/ 间质性膀胱炎（BPS/IC）的临 床疗效。方法 将63 例BPS/IC 患者按随机单盲的原则分成A、B 两组,A 组32 例,B 组31 例。A 组患者予膀胱灌注透明质酸钠40ml;B 组予膀胱灌注碱化利多卡因联合肝素治疗,即用5%碳酸氢钠10ml、2%利多卡因10ml、肝素钠2.5 万U 和0.9%氯化钠15ml的混合液行膀胱灌注。两组均保留40~60min,每周2 次,4周后改为每周1 次,共16 次。观察两组患者用药前及用药开始后4、12周时的O’Leary-Sant平均症状评分（ICPI）、问题评分（ICSI）及生活质量评分（QOL）,并记录不良反应。结果 两组患者治疗开始后4周和12 周ICPI、ICSI 及QOL评分均有明显改善,与治疗前比较均有统计学差异（均P＜0.05）。治疗4 周后两组的ICPI、ICSI 及QOL 评分下降分值比较均无统计学差异（均P＞0.05）,但治疗12 周后A 组的ICPI、ICSI 及QOL评分下降分值较B 组更为明显（均P＜0.05）。所有患者治疗过程中均未发生明显不良反应。结论 透明质酸钠及碱化利多卡因联合肝素膀胱灌注均能改善BPS/IC患者的临床症状,而透明质酸钠疗效更为显著。     

Highly sensitive and rapid determination of tacrolimus in peripheral blood mononuclear cells by liquid chromatography–tandem mass spectrometry

After solid organ transplantation, tacrolimus is given to prevent rejection. Therapeutic drug monitoring is used to reach target concentrations of tacrolimus in whole blood. Because the site of action of tacrolimus is the lymphocyte, and tacrolimus binds ~80% to erythrocytes, the intracellular tacrolimus concentration in lymphocytes is possibly more relevant. For this purpose, we aimed to develop, improve and validate a UPLC–MS/MS method to measure tacrolimus concentrations in isolated peripheral blood mononuclear cells (PBMCs). PBMCs were isolated using a Ficoll separation technique, followed by a washing step using red blood cell lysis. A cell suspension of 50 μL containing 1 million PBMCs was used in combination with MagSiMUS‐TDM^PREP. To each sample we added 30 μL lysis buffer, 20 μL reconstitution buffer containing ¹³C²H₄‐tacrolimus as internal standard, 40 μL MagSiMUS‐TDM^PREP Type I Particle Mix and 175 μL Organic Precipitation Reagent VI for methanol‐based protein precipitation. A 10 μL aliquot of the supernatant was injected into the UPLC–MS/MS system. The method was validated, resulting in high sensitivity and specificity. The method was linear (r² = 0.997) over the range 5.0–1250 pg/1 × 10⁶ PBMCs. The inaccuracy was <5% and the imprecision was <15%. The washing steps following Ficoll isolation could be performed at either room temperature or on ice, with no effect of the temperature on the results. A method for the analysis of tacrolimus concentrations in PBMCs was developed and successfully validated. Further research will be performed to investigate the correlation between concentrations in PBMCs and clinical outcome.     

Metabolomics study on promoting blood circulation and ameliorating blood stasis: Investigating the mechanism of Angelica sinensis and its processed products

Angelica sinensis (Danggui, DG) parched with alcohol (Jiu Danggui, JDG) and charred DG are the main processed products of DG, which are used to treat blood stasis syndrome (BSS). However, their therapeutic effect and mechanisms are still unclear. Based on an acute rat BSS model, the intervention effects of DG and its processed products (DGPPs) were evaluated by the hemorheology and coagulation function parameters. Meanwhile, plasma and urine metabolites were detected and analyzed by liquid chromatography coupled to quadrupole time‐of‐flight mass spectrometry and multivariate statistical analysis method. The results of hemorheology, coagulation function parameters and metabolomics all showed that the BSS model was successfully established, DGPPs intervention could significantly relieve rats BSS and the therapeutic effect of JDG was best. Moreover, 23 differential metabolites (14 in plasma and nine in urine) were identified that were closely related to the BSS, involving seven potential target metabolic pathways. DGPP intervention showed different degrees of reverse effect on these metabolites. JDG was the most effective owing to extensive regulation effect on differential metabolites. This study provides a reference for understanding the pathological mechanism of BSS and the mechanism of DGPPs, which lays a theoretical foundation for the rational use of DGPPs in clinical practice.     

Analysis of carbenoxolone by ultra‐high‐performance liquid chromatography tandem mass spectrometry in mouse brain and blood after systemic administration

Carbenoxolone is a derivative of glycyrrhetinic acid found in the root of Glycyrrhiza glabra, colloquially known as licorice. It has been used as a treatment for peptic and oral ulcers. In recent years, carbenoxolone has been utilized in basic research for its ability to block gap junctional communication. Better understanding the distribution of carbenoxolone after systemic administration can lead to a better understanding of its potential sites of action. Presented is an ultra high‐performance liquid chromatography tandem mass spectrometer (UHPLC–MS/MS) method for the identification and quantification of carbenoxolone in mouse blood and brain tissue. Twenty mice were injected intraperitoneally with 25 mg/kg carbenoxolone and brain tissue and blood were collected for analysis. Blood concentrations (mean ± SD) at 15, 30, 60 and 120 min were determined to be (n = 5) 5394 ± 778, 2636 ± 836, 1564 ± 541 and 846 ± 252 ng/mL, respectively. Brain concentrations (mean ± SD) at 15, 30, 60 and 120 mins were determined to be (n = 5) 171 ± 62, 102 ± 35, 55 ± 10 and 27 ± 9 ng/g, respectively. The analysis of these specimens at the four different time points resulted in blood and brain half‐lives in mice of ~43 and 41 min, respectively. The UHPLC–MS/MS method was determined to be sensitive and robust for quantification of carbenoxolone.     

Quantitative analysis of enasidenib in dried blood spots of mouse blood using an increased‐sensitivity LC–MS/MS method: Application to a pharmacokinetic study

A simple, sensitive and rapid assay method has been developed and validated as per regulatory guidelines for the estimation of enasidenib on mouse dried blood spots (DBS) using liquid chromatography coupled to tandem mass spectrometry with electrospray ionization in the positive‐ion mode. The method employs liquid extraction of enasidenib from DBS disks of mouse whole blood followed by chromatographic separation using 0.2% formic acid–acetonitrile (25:75, v/v) at a flow rate of 1.0 mL/min on an Atlantis dC₁₈ column with a total run time of 2.0 min. The MS/MS ion transitions monitored were m/z 474.0 → 267.1 for enasidenib and m/z 309.2 → 251.3 for the internal standard (warfarin). The assay was linear in the range of 1.01 – 3044 ng/mL. The within‐run and between‐run precisions were in the range of 3.18 – 9.06 and 4.66 – 8.69%, respectively. Stability studies showed that enasidenib was stable on DBS cards for 1 month. This novel method has been applied to analyze the DBS samples of enasidenib obtained from a pharmacokinetic study in mice.     

Validation of a HPLC/MS method for simultaneous quantification of clonidine,morphine and its metabolites in human plasma

A high‐performance liquid chromatography–tandem mass spectrometry method was developed and validated for the simultaneous quantification of morphine, morphine's major metabolites morphine‐3‐glucuronide and morphine‐6‐glucuronide, and clonidine, to support the pharmacokinetic analysis of an ongoing double‐blinded randomized clinical trial that compares the use of morphine and clonidine in infants diagnosed with neonatal abstinence syndrome. Plasma samples were processed by solid‐phase extraction and separated on an Inertsil ODS‐3 (4 μm) column using an 0.1% formic acid in water–0.1% formic acid in methanol gradient. Detection of the analytes was conducted in the positive multiple reaction monitoring mode. The range of quantitation was 1–1000 ng/mL for morphine, morphine‐3‐glucuronide and morphine‐6‐glucuronide, and 0.25–100 ng/mL for clonidine. Intra‐day and inter‐day accuracy and precision were ≤15% for all analytes across the quantitation range. Extraction recovery rates were ≥94% for morphine, ≥90% for M3G, ≥87% for M6G and ≥ 79% for clonidine. Matrix effect ranged from 85–94% for clonidine to 101–106% for M3G. The method fulfilled all predetermined acceptance criteria and required only 100 μL of starting plasma volume. Furthermore, it was successfully applied to 30 clinical trial plasma samples.     

Screening of anti-inflammatory and antioxidant potential of functionalized tetrahydrocarbazole linked 1,2-diazoles and their docking studies

Inflammatory diseases are associated with life-threatening syndromes like hepatitis, cancer, and trauma injury while some decrease the quality of life such as rheumatism, arthritis, and tuberculosis. 1,2-Diazoles (pyrazolines) play a vital role in COX-2 inhibition thus dinitro-tetrahydrocarbazole linked pyrazolines have been synthesized and endeavor to screen for anti-inflammatory, antioxidant and molecular docking studies. For this purpose, 6,8-dinitro-acetyl-2,3,4,9-tetrahydrocarbazole (I), aromatic aldehydes (IIa-e) and hydrazines (IIIa-b) were combined via multicomponent reaction approach under the influence of microwave irradiations to afford pyrazolines (1–10). All new molecules were screened for in vitro anti-inflammatory activity by human red blood cells membrane stabilization, antioxidant potential by2,2-diphenyl-1-picrylhydrazyl,2,2´-azinobis (3-ethylbenzo thiazoline)-6-sulphonic acid, lipid peroxidation, and total antioxidant capacity assays along with cytotoxicity by brine shrimp lethality assay. Molecular docking was performed by using the Auto Dock program. Both disubstituted and trisubstituted diazoles showed excellent membrane stabilizing effects, (91.89 % and 77 %, respectively). The presence of phenol, furan, thiocarbamide, and chloro-moieties have the most prominent effect. Toxicity results indicated that compounds were less toxic at the tested dose (0.1 mg/ml). The antioxidant study showed that compound 2 was more active showing low IC₅₀ values (32.2 and 39.2 µg/ml) in DPPH and total phenolic contents assays respectively. Compound 3 (44.0 µg/ml) showed the highest potential assay in ABTS radical neutralization assay while compound 7 (65.0 µg/ml) showed maximum potential in lipid peroxidation. All diazoles (1–10) were screened for in vitro anti-inflammatory potential where disubstituted diazoles were found better than trisubstituted analogs and exhibited significant antioxidant potential. Molecular docking of diazoles showed a good correlation of their anti-inflammatory activity with p38α MAPK, COX-2, and 5-LOX enzymes that are molecular therapeutic targets of inflammation.     

Cell Membrane-camouflaged Multi-functional Dendritic Large Pore Mesoporous Silica Nanoparticles for Combined Photothermal Therapy and Radiotherapy of Cancer

Combining photothermal therapy and radiotherapy(PTT-RT) with reducing tumor hypoxia acts as an important antitumor modality. However, it is a great challenge to realize photothermal therapy, radiotherapy and exogenous oxygen supply in one nanosystem. To realize a combination of the three functions, we fabricated a red blood cell membrane(RBCm)-camouflaged, red blood cell content(RBCc) and the copper sulfide(CuS) co-loaded dendritic large pore mesoporous silica nanoparticle(DLMSN/CuS/RBCc/ RBCm). The cell membrane coating endowed the nanoparticles with good stability in the physiological environment, and CuS allowed the nanoparticle exhibiting good photothermal and radiosensitization properties. RBCc loaded nanoparticle DLMSN/CuS/RBCc enhanced superior anti-tumor effect than DLMSN/CuS during combined PTT-RT therapy because the introduction of RBCc increased the exogenous oxygen supply. The in vitro study further demonstrated that the combination of photothermal therapy and radiotherapy induced superior antitumor efficacy than single therapy. Our work thus presents a unique multifunctional nanoscale platform favorable for combined PTT and RT.     

Tetrachloroferrate(III)-selective liquid membrane electrode based on crystal violet

An extract of crystal violet-tetrachloroferrate(III) in nitrobenzene was used to prepare a tetrachloroferrate(III)-selective liquid membrane electrode with a poly(vinyl chloride) support. The optimal conditions to determine 2.5 × 10^–5 – 5.0 × 10^–2 M iron(III) as tetrachloroferrate(III) (anionic slope 56 mV/decade, detection limit 7.9 × 10^–6 M) were found to be 4.0–5.5.M total chloride in 0.75–1.5M hydrochloric acid. The electrode was reliably applied to determine iron in human blood, haematite and mineralized vitamin syrup by direct potentiometry, standard and sample additions as well as standard subtraction techniques.     

Down syndrome critical region 1 enhances the proteolytic cleavage of calcineurin

Down syndrome critical region 1 (DSCR1), an oxidative stress-response gene, interacts with calcineurin and represses its phosphatase activity. Recently it was shown that hydrogen peroxide inactivates calcineurin by proteolytic cleavage. Based on these facts, we investigated whether oxidative stress affects DSCR1-mediated inactivation of calcineurin. We determined that overexpression of DSCR1 leads to increased proteolytic cleavage of calcineurin. Convertsely, knockdown of DSCR1 abolished calcineurin cleavage upon treatment with hydrogen peroxide. The PXIIXT motif in the COOH-terminus of DSCR1 is responsible for both binding and cleavage of calcineurin. The knockdown of overexpressed DSCR1 in DS fibroblast cells also abrogated calcineurin proteolysis by hydrogen peroxide. These results suggest that DSCR1 has the ability to inactivate calcineurin by inducing proteolytic cleavage of calcineurin upon oxidative stress.