Simultaneous Determination of Six Compounds in Rat Plasma by Ultra‑Performance Liquid Chromatography with Tandem Mass Spectrometry: Application in the Pharmacokinetic Study of Qing Gan‑Shu Yu‑Fang

A rapid and high selective ultra?performance liquid chromatography (UPLC) with tandem mass spectrometry method for simultaneous determination of six compounds including albiflorin, paeoniflorin, picroside I, picroside II, saikosaponin A, and saikosaponin D in rat plasma was developed and validated using butyl p-hydroxybenzoate as an internal standard. One-step direct protein precipitation with acetonitrile was used to extract the compounds from the rat plasma samples. Chromatographic separation was achieved using an ACQUITY UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 µm) at a flow rate of 0.4 mL/min, using gradient mode containing 0.1% formic acid in water and acetonitrile were used as the Mobile phase A and B. Electrospray ionization in negative ion mode and multiple reaction monitoring were used to identify and quantify active components. Calibration curves showed good linearity (R2 > 0.9908) over a wide concentration range for all compounds. The intra- and interday precision (relative standard deviation) ranged 2.4%–7.0% and 2.6%–8.0%, respectively. The accuracy (relative error) was from ?13.0% to 13.2% at all quality control levels. The recovery ranged from 81.1% to 92.5%. The validated method was successfully applied to pharmacokinetic study in rats after oral administration of Qing Gan?Shu Yu?Fang. The results show that one can draw a conclusion that these six active ingredients can be quickly absorbed and play a pharmacodynamic role rapidly in vivo.     

Advantages of Anti-Inflammatory Acupuncture in Treating Sepsis of Coronavirus Disease 2019

Background: Sepsisis one of the mostserious complications and a leading cause of death in patients with coronavirus disease 2019 (COVID-19). In general, it isthe result of an unregulated inflammatory cascade such as a postinfection “cytokine storm.” The conventional treatment mainly relies on glucocorticoids, of which curative effects are not ideal, as they come with significant side effects. It is critical to seek or develop other effective therapeutics in dealing cytokine storm to fight COVID-19 with sepsis. Aims and Objectives: Raise awareness of the significance applying anti-inflammatory acupuncture in dealing COVID-19 patients with sepsis and provide an appropriate acupuncture protocol that can be easily integrated into existing medical guideline. Materials and Methods: Current evidences from animal experiments and clinical trials about acupuncture in treating infectious sepsis are reviewed, and a detailed discussion on advantages of anti?inflammatory acupuncture is followed, then the rationality on the point selection and stimulation parameters of acupuncture is analyzed to propose an appropriate acupuncture protocol. Results: Current experiments have shown that acupuncture can play a significant role to improve inflammation reaction and reduce mortality in infectious animal and patients with sepsis and its mechanisms are mainly achieved by stimulating the vagus?cholinergic anti?inflammatory pathways.Applying acupuncture in treating COVID-19 patients with sepsis hasfour aspects of advantages. Moreover, a simple and convenient clinical acupuncture protocol including point selection and appropriate stimulation parameters is proposed. Conclusion: Acupuncture, especially electroacupuncture, has shown potentials in effectively treating infectious sepsis of animal models and critically ill patients in small sample studies by stimulating the nervous system, but has been largely overlooked in the clinic so far. It is advised that acupuncture should be integrated into the existing medical guidelines in dealing with COVID-19 complicated with sepsis.     

Antiviral Properties of Traditional Chinese Medicine against Coronavirus: Research Clues for Coronavirus Disease-2019

The objective of this study was to provide research clues for the prevention and treatment of coronavirus disease?2019 (COVID?19) and coronavirus (CoV) infection using Traditional Chinese Medicine (TCM). A review on research and clinical trials that using TCM extracts and active ingredients against CoV was performed, and a table of TCM agents and their effects on CoV were summarized. Relevant analysis was performed and visual expression of the data included summarizing the types of TCM and treatment methods for COVID?19. TCM fighting against CoV is mainly used in the lung and heart channels, and its medicinal properties are mainly cold and mild, while its taste is mainly bitter and sweet. The majority of research focused on treatments that clear away heat and toxic materials and those that strengthen body resistance and tonify deficiencies. TCM has unique advantages to fight against CoV. The development of new anti?CoV therapy using TCM is of great significance for the prevention and treatment of COVID?19 pneumonia and various viral infectious diseases.     

从阴阳平调试论《傅青主女科》治疗妇人痛证

从阴阳平调的角度探究《傅青主女科》治疗妇人痛证,分析傅青主运用阴阳互用、互制理论,治疗妇人经行腹痛、妊娠腹痛及产后诸痛的特点。傅青主治疗妇人痛证以阴阳平和为本,以气血相和治疗经水不利而腹痛,以水火相济治疗经前腹痛与妊娠躁狂,以动静相合治疗产后腹痛,以寒热合宜治疗产后心、腹痛证。书中记载傅青主论治妇人痛证时,补气兼有养血,泻火亦护阴液,散寒且温阳,益气血又行气血,用药纯和,重视炮制,少用贵重峻烈之药,以阴阳平调为期,体现了傅青主重视阴阳,善用阴阳平调的治疗理念。     

Ultra-High Performance Liquid Chromatography/Ion Mobility-Quadrupole Time-of-Flight Mass Spectrometry and Database-Driven Automatic Peak Annotation for the Rapid Profiling and Characterization of the Multicomponents from Stephaniae Tetrandrae Radix (Fang-Ji)

Quality control of traditional Chinese medicine (TCM) begins with elucidation of its chemical basis. The root of Stephania tetrandra, Stephaniae Tetrandrae radix (STR; Fang-Ji), has long been utilized as an antirheumatic, analgesic, and diuretic TCM. Powerful analytical strategies that would enable a multicomponent characterization of STR are still lacking. In the present study, we established a rapid, reliable, and enhanced profiling approach, using ultra?high performance liquid chromatography coupled with ion mobility/quadrupole time?of?flight mass spectrometry (MS) and automatic peak annotation facilitated by computational matching of an in?house library. This approach was used to characterize the multicomponents of STR. Good chromatographic separation was achieved within 17 min on a reversed-phase BEH C18 column eluted with acetonitrile/0.1% ammonium hydroxide in water, whereas data?independent high?definition MSE in the positive mode was applied to acquire the MS2 data using a Vion? IM?QTOF instrument, which, in theory, could cover all the profiled precursor ions. To overcome the interference of three predominant peaks, a knockout strategy was utilized by automated valve switching. An in?house library of 163 compounds was established and incorporated into the UNIFI? platform. By applying this method, we could identify or tentatively characterize 76 alkaloidsfrom the methanolic extract of STR, including 14 aporphine?type, four morphine?type, 48 bisbenzylisoquinoline?type, seven tetrahydroprotoberberine?type, one protopine?type, one benzylisoquinoline?type, and one other. For each component, four?dimensional information, such as retention time, collision cross-section, high-accuracy MS1, and high-accuracy MS2 data, was utilized to achieve the systematic multicomponent characterization of STR.     

Simultaneous Determination and Pharmacokinetics of Tetrandrine, Fangchinoline, and Cyclanoline in Rat Plasma by Ultra-High Performance Liquid Chromatography-Mass Spectrometry after Oral Administration of Stephaniae Tetrandrae Radix Extract

Objective: The objective of the study wasto develop a rapid and sensitive ultra?performance liquid chromatography?tandem massspectrometric method for the determination of tetrandrine, fangchinoline, and cyclanoline in rat plasma and to investigate their pharmacokinetics after oral administration of Stephaniae Tetrandrae Radix extracts. Methods: Sample pretreatment involved methanol pretreatment and liquid–liquid extraction of ethyl acetate from plasma with methanol. Tramadol was used as the internal standard. The analysis was performed using an high strength silica T3 column (100 mm × 2.1 mm, 1.8 μm) and a gradient elution method consisting of mobile phase solution A (0.1% formic acid in water) and B (acetonitrile) at a flow rate of 0.4 mL/min. The detection was performed using a triple quadrupole tandem mass spectrometer in the multiple reaction monitoring mode and using an electrospray ionization source in the positive ionization mode. Results: High efficiency was achieved with an analysis time of 4 min/sample. The calibration curve linear in the concentration range of 1250 ng/ml (R2 ≥ 0.9900) and the lower limit of quantification is 1 ng/ml. The intraday and interday precision (relative standard deviation) values were lower than 9.4. Accuracy (relative error) was within 10.3% at all three quality control levels. Conclusions: This method was successfully applied in pharmacokinetics of tetrandrine, fangchinoline, and cyclanoline in rats after oral administration of Stephaniae Tetrandrae Radix extracts. The maximum plasma concentration (Cmax) of tetrandrine, fangchinoline, and cyclanoline was 124.71 ± 16.08, 84.56 ± 3.28, and 57.61 ± 6.26 ng/ mL, respectively. The time to reach Cmax was 10.39 ± 3.04 for tetrandrine, 10.17 ± 3.04 for fangchinoline, and 6.40 ± 3.16 for cyclanoline. The pharmacokinetic results might help further guide the clinical application of Stephaniae Tetrandrae Radix.     

MicroRNA and mRNA profiling of cerebral cortex in a transgenic mouse model of Alzheimer’s disease by RNA sequencing

In a previous study,we found that long non-coding genes in Alzheimer’s disease(AD)are a result of endogenous gene disorders caused by the recruitment of microRNA(miRNA)and mRNA,and that miR-200a-3p and other representative miRNAs can mediate cognitive impairment and thus serve as new biomarkers for AD.In this study,we investigated the abnormal expression of miRNA and mRNA and the pathogenesis of AD at the epigenetic level.To this aim,we performed RNA sequencing and an integrative analysis of the cerebral cortex of the widely used amyloid precursor protein and presenilin-1 double transgenic mouse model of AD.Overall,129 mRNAs and 68 miRNAs were aberrantly expressed.Among these,eight down-regulated miRNAs and seven up-regulated miRNAs appeared as promising noninvasive biomarkers and therapeutic targets.The main enriched signaling pathways involved mitogen-activated kinase protein,phosphatidylinositol 3-kinase-protein kinase B,mechanistic target of rapamycin kinase,forkhead box O,and autophagy.An miRNA-mRNA network between dysregulated miRNAs and corresponding target genes connected with AD progression was also constructed.These miRNAs and mRNAs are potential biomarkers and therapeutic targets for new treatment strategies,early diagnosis,and prevention of AD.The present results provide a novel perspective on the role of miRNAs and mRNAs in AD.This study was approved by the Experimental Animal Care and Use Committee of Institute of Medicinal Biotechnology of Beijing,China(approval No.IMB-201909-D6)on September 6,2019.     

瘢痕子宫再次妊娠产妇行椎管内分娩镇痛的效果

目的观察瘢痕子宫再次妊娠产妇行椎管内分娩镇痛的安全性和有效性。方法选择2017年5月至2018年4月我院收治的瘢痕子宫再次妊娠同意阴道试产单胎足月头位产妇101例,随机选取分娩镇痛产妇70例为观察组,其中硬膜外分娩镇痛(E组)36例,腰-硬联合分娩镇痛(C组)34例,同期未镇痛产妇31例为对照组(N组)。记录产妇各产程时间、出血量、新生儿1、5 min Apgar评分、阴道分娩、产钳助产、子宫破裂情况,记录镇痛前、给药后5、10、20 min产妇NRS评分和Bromage评分。结果与N组比较,E组和C组第二产程时间明显延长(P0.05);E组出血量高于N组及C组,但差异无统计学意义。三组第一、第三产程时间、新生儿1、5 min Apgar评分、阴道分娩、产钳助产、子宫破裂发生率差异无统计学意义;镇痛后E组和C组NRS评分均呈下降趋势。与E组比较,C组NRS评分明显降低(P0.05)。结论瘢痕子宫再次妊娠产妇采用椎管内分娩镇痛安全可行,不降低阴道分娩率,不增加出血量以及产钳助产、子宫破裂发生率。腰-硬联合分娩镇痛较硬膜外分娩镇痛的镇痛效果好。     

A Network Pharmacology Study of Reduning Injection for the Treatment of Coronavirus Disease-19

Objective: The 2019 novel coronavirus (2019?nCoV) outbreak has escalated into a global pandemic. According to Chinese guidance for coronavirus disease?19 (COVID?19): Prevention, control, diagnosis, and management, Reduning injection can effectively treat, the disease caused by the virus. To identify the active ingredients of Reduning injection and COVID?19 disease?related pathways, we conducted a network pharmacology study. Methods: The Traditional Chinese Medicine Systems Pharmacology database was used to screen the chemical constituents and potential targets of Reduning injection. The gene names were converted to the corresponding protein names using UniProt. GeneCards and OMIM databases were used to select targets related to 2019?nCoV. Using Cytoscape 3.7.2 software platform and STRING database, we constructed drug?common target and target protein protein?protein interaction network diagrams. Rx64 3.6.2 software and Bioconductor biological information software package were used for? Gene Ontology (GO) functional enrichment and KEGG pathway analyses. Results: In Reduning injection, a total of 33 effective chemical components were obtained that were involved in 151 signaling pathways, of which 44 targets were considered therapeutically relevant. Conclusion: Reduning injection can be potentially applied for the treatment of COVID-19 based on the results of our network pharmacology study.