Inhibition of total flavonoids from Abelmoschus Manihot on cytochrome P450 / 中国药科大学学报

@#To investigate the influential mechanism of total flavonoids from Abelmoschus Manihot (HKZ) on cytochrome P450 (CYP450) isoforms in human liver microsomes and to verify its effect on the most significantly inhibited subtype CYP2C9 in rats.The inhibitory effects of HKZ on human CYP3A4, CYP2C9, CYP2C19, CYP2E1, CYP1A2 and CYP2D6 were evaluated through the cocktail method using ultra-performance liquid chromatography tandem mass spectrometry, then its inhibitory mechanism was investigated and kinetic parameters of enzyme inhibition were calculated By comparing the pharmacokinetic behaviors of tolbutamide after single or multiple administration of 200 mg/kg HKZ and equal dose of CMC-Na in rats, the effects of HKZ on CYP2C11 enzyme (CYP2C9 isoenzyme) was estimated.The results indicated the significant inhibitory effect of HKZ on CYP2C9 and CYP2E1 with IC50 of 3.22 and 8.64 μg/mL, respectively. Also, it showed certain inhibitory ability on other isoforms with IC50 of 20-30 μg/mL.As demonstrated, HKZ may not be a time-dependent inhibitor which competitively inhibited CYP2E1 and CYP2C9 with Ki of 3.84 and 6.33 μg/mL.In contrast, it showed noncompetitive inhibition on CYP3A4 mediated testosterone-6β-hydroxylation and midazolam-4-hydroxylation reaction with Ki of 7.37 and 3.32 μg/mL.It was also a noncompetitive inhibitor of CYP1A2, CYP2D6 and CYPC219 with Ki values of 8.66, 11.49 and 21.94 μg/mL. HKZ did not change the pharmacokinetic parameters of CYP2C11 probe substrate tolbutamide in rat, but it affected the AUC0-t, cmax of 4-hydroxytolubutamide (P < 0.05). Therefore, drug-drug interaction mediated by CYP450 should be considered in clinical study.

The role of phosphatidylcholine 34:1 in the occurrence, development and treatment of ulcerative colitis

Lipid homeostasis is considered to be related to intestinal metabolic balance, while its role in the pathogenesis and treatment of ulcerative colitis (UC) remains largely unexplored. The present study aimed to identify the target lipids related to the occurrence, development and treatment of UC by comparing the lipidomics of UC patients, mice and colonic organoids with the corresponding healthy controls. Here, multi-dimensional lipidomics based on LC-QTOF/MS, LC-MS/MS and iMScope systems were constructed and used to decipher the alteration of lipidomic profiles. The results indicated that UC patients and mice were often accompanied by dysregulation of lipid homeostasis, in which triglycerides and phosphatidylcholines were significantly reduced. Notably, phosphatidylcholine 34:1 (PC34:1) was characterized by high abundance and closely correlation with UC disease. Our results also revealed that down-regulation of PC synthase PCYT1α and Pemt caused by UC modeling was the main factor leading to the reduction of PC34:1, and exogenous PC34:1 could greatly enhance the fumarate level via inhibiting the transformation of glutamate to N-acetylglutamate, thus exerting an anti-UC effect. Collectively, our study not only supplies common technologies and strategies for exploring lipid metabolism in mammals, but also provides opportunities for the discovery of therapeutic agents and biomarkers of UC.

SIRT1 activation synergizes with FXR agonism in hepatoprotection via governing nucleocytoplasmic shuttling and degradation of FXR

Farnesoid X receptor (FXR) is widely accepted as a promising target for various liver diseases; however, panels of ligands in drug development show limited clinical benefits, without a clear mechanism. Here, we reveal that acetylation initiates and orchestrates FXR nucleocytoplasmic shuttling and then enhances degradation by the cytosolic E3 ligase CHIP under conditions of liver injury, which represents the major culprit that limits the clinical benefits of FXR agonists against liver diseases. Upon inflammatory and apoptotic stimulation, enhanced FXR acetylation at K217, closed to the nuclear location signal, blocks its recognition by importin KPNA3, thereby preventing its nuclear import. Concomitantly, reduced phosphorylation at T442 within the nuclear export signals promotes its recognition by exportin CRM1, and thereby facilitating FXR export to the cytosol. Acetylation governs nucleocytoplasmic shuttling of FXR, resulting in enhanced cytosolic retention of FXR that is amenable to degradation by CHIP. SIRT1 activators reduce FXR acetylation and prevent its cytosolic degradation. More importantly, SIRT1 activators synergize with FXR agonists in combating acute and chronic liver injuries. In conclusion, these findings innovate a promising strategy to develop therapeutics against liver diseases by combining SIRT1 activators and FXR agonists.

Quantitative determination of endogenous histamine and histidine in biological matrices by double adsorption based on HPLC-MS/MS / 中国药科大学学报

@#An innovative approach to quantitatively analyze the histamine and its precursor histidine simultaneously in biological matrices was established for the first time based on double adsorption combined with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).The internal standard was 2-dihydroxybenzoic acid (DHB).The plasma and brain tissue homogenate was protein precipitated with 3-fold acetonitrile, and the supernatant was then sampled for injection analysis.The chromatographic separation of the target components was achieved on an amino chromatography column (ODS-SPXBridge? Amide).Gradient elution was carried out with the mobile phase consisting of solvent A (0.1% formic acid and 1mmol/L ammonium formate in water) and solvent B (acetonitrile).Mass spectrometry was employed for quantitative analysis with ESI ion source in multiple reaction monitoring (MRM) mode.In order to improve the specificity and accuracy, activated carbon and calcite were used for the double adsorption of biological matrices for the first time.The adsorbed matrix was then used for methodology validation.The results showed that histamine and histidine were linear in the quantitative range (correlation coefficient r ≥ 0.999).Accuracy, precision, extraction recovery, matrix effect and stability all met the requirements of biological sample analysis.All results suggested that the present method could not only be efficiently and reliably used for simultaneous quantitative analysis of histamine and histidine in biological samples, but also provide reference for the detection of other endogenous substances.

Pharmacokinetic study of Erlong Zuoci Pill in rats / 中国药科大学学报

@#To establish a quantitative LC-MS/MS method for the simultaneous detection of components of Erlong Zuoci Pill in rat plasma: verbascoside, oxypaeoniflorin, echinacoside and benzoylpaeoniflorin, and to evaluate the pharmacokinetic characteristics of Erlong Zuoci Pill in rats, plasma samples were purified by protein precipitation using methanol as a protein precipitant.Methanol was used as the organic phase and aqueous solution containing 0.1% formic acid was used as the water phase.The quantitative analysis method of verbascoside, oxypaeoniflorin, echinacoside and benzoylpaeoniflorin was established in negative ion mode, and the validation of bioanalytical method was carried out.Healthy SD rats were selected, and 20 mL/kg (equivalent to the original drug 10 g/kg dose) of Erlong Zuoci Pill extract was administered by intragastric administration.The plasma concentration of the target compounds at different time intervals after administration was determined using the established method, and the pharmacokinetic parameters was calculated by the Phoenix WinNonlin8.3 software using the non-compartmental model.The method validation results showed that verbascoside (r = 0.993 7) and oxypaeoniflorin (r = 0.994 6) had good linear relationship in the concentration range of 0.5-50 ng/mL, echinacoside (r = 0.993 6) and benzoylpaeoniflorin (r = 0.992 6) had good linear relationship in the concentration range of 1-100 ng/mL.The relative standard deviations of the inter- and intra- batch precision of the four compounds were all less than 15%, and the inter- batch and intra- accuracies were between 85% and 115%.Extraction recovery, matrix effect and stability met the relevant requirements.After a single gavage of Erlong Zuoci Pill extract in rats, all the four compounds were rapidly absorbed and eliminated.Oxypaeoniflorin, echinacoside, and benzoylpaeoniflorin showed two peaks in their drug concentration-time curves.Compared with the other three compounds, oxypaeoniflorin has the highest concentration in rat plasma with cmax1 of (24.40 ± 4.78) ng/mL and cmax2 of (22.50 ± 2.70) ng/mL. The results show that the validation results of this method are in line with the guiding principles of biological sample analysis methods, and it can be used to evaluate the pharmacokinetic characteristics of Erlong Zuoci Pill extract in rats.

Advances in research on visualization of living cell drugs in vivo / 中国药科大学学报

@#The development of living cell drugs and their successful application in clinical treatments require full clarification of the fate of cells after transplantation, which is critical to the safety and efficacy of living cell drugs.In order to solve this problem, cell imaging technology has come into our sight, and the use of visualization technology for non-invasive tracing of living cell drugs could reveal the distribution, homing and activity of living cell drugs in the body, which helps to determine the best number of transplanted cells, optimize the administration scheme, improve the transplantation efficiency, enhance the targeting of transplanted cells, and reduce the potential off-target accumulation risk.This paper summarizes the research advances of non-invasive visual tracing in vivo for living cell drugs from the perspectives of radionuclide imaging, magnetic resonance imaging, magnetic particle imaging, computed tomography imaging, fluorescence imaging and multimodal imaging.The aim is to obtain the biological behavior of living cell drugs in vivo with the application of appropriate contrast agent and tracing technology, and provide a more reasonable scientific basis for the research and development of living cell drugs and their transplantation therapy.

Mechanism of sodium salicylate-induced damage to HEI-OC1 cells based on metabonomics / 中国药科大学学报

@#The effect of sodium salicylate on the endogenous metabolism of hair cell-like cells (HEI-OC1).of mice was analyzed based on liquid chromatography-quadrupole time of flight mass spectrometry (LC-Q-TOF/MS).Firstly, HEI-OC1 cells were treated with different concentrations of sodium salicylate, and cell survival was examined by the CCK-8 method. Next, sodium salicylate was administered for different duration to observe the changes in cell morphology. Inter-group differential metabolites were screened out, and the associated metabolic pathways were analyzed based on metabonomic technology.Results showed that sodium salicylate could significantly inhibit the survival rate of HEI-OC1 cells, and that, as the concentration increased, the inhibitory effect became stronger. Also, the cell morphology could be elongated after administration and return to normal after withdrawal.Eighteen differential metabolites such as orotic acid, uridine and aspartic acid were screened out after treatment of sodium salicylate, which mainly involving two possible metabolic pathways, namely the metabolism of alanine, aspartic acid and glutamic acid, and that of pyrimidine.In summary, the application of metabolomics technology to evaluate the effect of sodium salicylate on hair cells from the microscopic perspective can provide new ideas for the study of sodium salicylate ototoxicity and development of tinnitus.

Target-responsive subcellular catabolism analysis for early-stage antibody-drug conjugates screening and assessment

Events including antibody‒antigen affinity, internalization, trafficking and lysosomal proteolysis combinatorially determine the efficiency of antibody-drug conjugate (ADC) catabolism and hence the toxicity. Nevertheless, an approach that conveniently identifies proteins requisite for payload release and the ensuing toxicity for mechanistic studies and quality assessment is lacking. Considering the plethora of ADC candidates under development, we developed a target-responsive subcellular catabolism (TARSC) approach that examines ADC catabolism and probes changes in response to targeted interferences of proteins of interest. We firstly applied TARSC to study the commercial T-DM1 and the biosimilar. We recorded unequivocal catabolic behaviors regardless of the absence and presence of the targeted interferences. Their negligible differences in TARSC profiles agreed with their undifferentiated anti-tumoral efficacy according to further

Comparative analysis of constitutes and metabolites for traditional Chinese medicine using IDA and SWATH data acquisition modes on LC-Q-TOF MS / 药物分析学报

Identification of components and metabolites of traditional Chinese medicines (TCMs) employing liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF MS) techniques with information-dependent acquisition (IDA) approaches is increasingly frequent. A current drawback of IDA-MS is that the complexity of a sample might prevent important compounds from being triggered in IDA settings. Sequential window acquisition of all theoretical fragment-ion spectra (SWATH) is a data-independent acquisition (DIA) method where the instrument deterministically fragments all precursor ions within the predefined m/z range in a systematic and unbiased fashion. Herein, the superiority of SWATH on the detection of TCMs' components was firstly investigated by comparing the detection ef-ficiency of SWATH-MS and IDA-MS data acquisition modes, and sanguisorbin extract was used as a mode TCM. After optimizing the setting parameters of SWATH, rolling collision energy (CE) and variable Q1 isolation windows were found to be more efficient for sanguisorbin identification than the fixed CE and fixed Q1 isolation window. More importantly, the qualitative efficiency of SWATH-MS on sanguisorbins was found significantly higher than that of IDA-MS data acquisition. In IDA mode, 18 kinds of sangui-sorbins were detected in sanguisorbin extract. A total of 47 sanguisorbins were detected when SWATH-MS was used under rolling CE and flexible Q1 isolation window modes. Besides, 26 metabolites of sangui-sorbins were identified in rat plasma, and their metabolic pathways could be deduced as decarbonylation, oxidization, reduction, methylation, and glucuronidation according to their fragmental ions acquired in SWATH-MS mode. Thus, SWATH-MS data acquisition could provide more comprehensive information for the component and metabolite identification for TCMs than IDA-MS.

Herbal drug discovery for the treatment of nonalcoholic fatty liver disease

Few medications are available for meeting the increasing disease burden of nonalcoholic fatty liver disease (NAFLD) and its progressive stage, nonalcoholic steatohepatitis (NASH). Traditional herbal medicines (THM) have been used for centuries to treat indigenous people with various symptoms but without clarified modern-defined disease types and mechanisms. In modern times, NAFLD was defined as a common chronic disease leading to more studies to understand NAFLD/NASH pathology and progression. THM have garnered increased attention for providing therapeutic candidates for treating NAFLD. In this review, a new model called "multiple organs-multiple hits" is proposed to explain mechanisms of NASH progression. Against this proposed model, the effects and mechanisms of the frequently-studied THM-yielded single anti-NAFLD drug candidates and multiple herb medicines are reviewed, among which silymarin and berberine are already under U.S. FDA-sanctioned phase 4 clinical studies. Furthermore, experimental designs for anti-NAFLD drug discovery from THM in treating NAFLD are discussed. The opportunities and challenges of reverse pharmacology and reverse pharmacokinetic concepts-guided strategies for THM modernization and its global recognition to treat NAFLD are highlighted. Increasing mechanistic evidence is being generated to support the beneficial role of THM in treating NAFLD and anti-NAFLD drug discovery.

Combined obeticholic acid and apoptosis inhibitor treatment alleviates liver fibrosis

Obeticholic acid (OCA), the first FXR-targeting drug, has been claimed effective in the therapy of liver fibrosis. However, recent clinical trials indicated that OCA might not be effective against liver fibrosis, possibly due to the lower dosage to reduce the incidence of the side-effect of pruritus. Here we propose a combinatory therapeutic strategy of OCA and apoptosis inhibitor for combating against liver fibrosis. CCl-injured mice, d-galactosamine/LPS (GalN/LPS)-treated mice and cycloheximide/TNF (CHX/TNF)-treated HepG2 cells were employed to assess the effects of OCA, or together with IDN-6556, an apoptosis inhibitor. OCA treatment significantly inhibited hepatic stellate cell (HSC) activation/proliferation and prevented fibrosis. Elevated bile acid (BA) levels and hepatocyte apoptosis triggered the activation and proliferation of HSCs. OCA treatment reduced BA levels but could not inhibit hepatocellular apoptosis. An enhanced anti-fibrotic effect was observed when OCA was co-administrated with IDN-6556. Our study demonstrated that OCA inhibits HSCs activation/proliferation partially by regulating BA homeostasis and thereby inhibiting activation of HSCs. The findings in this study suggest that combined use of apoptosis inhibitor and OCA at lower dosage represents a novel therapeutic strategy for liver fibrosis.

Advances in methodologies for predicting metabolic stability for low-clearance drugs / 中国药科大学学报

@#The metabolic stability test of drugs is a key step in drug discovery and achieving low clearance is frequently the goal in the design of drug. Increased drug metabolism stability can reduce drug dosage, enhance drug exposure and prolong drug half-life. Accurately assessing the metabolic stability parameters of low clearance drugs and predicting human pharmacokinetics has become a challenge. Traditional tools in vitro including microsomes and suspended primary hepatocytes are limited by incubation time, which is not long enough to make sufficient metabolic conversion. Determination of intrinsic clearance or metabolic pathways and mechanisms of drug are implicated. Novel models tend to further mimic the in vivo environment in order to prolong lifetime of hepatocytes and achieve sufficient metabolic turnover of drugs for monitoring. In vitro-in vivo correlation of intrinsic clearance of methodologies has evaluated to support the reliability in predicting human pharmacokinetics. Application of these methodologies greatly decreases the forthputting of experimental animals and the release of expensive clinical trials during the acquisition of pharmacokinetic parameters. In this review, we summarized the principles, advantages and disadvantages of the novel in vitro methodologies for metabolic stability dealing with low-turnover drugs, including hepatocyte relay method, plated human hepatocytes, coculture system and microfluidic devices. Future prospect is proposed for in vitro metabolic models and it provides reference and optimization in metabolic stability for early lead compounds.

Advances in molecular mechanisms of organelle interaction and their role in disease developmen / 中国药科大学学报

@#Organelles have their special functions, they interact with each other and coordinate a series of important physiological functions at the same time. Organelle interaction occurs at membrane contact sites(MCSs), where the membranous organelle endoplasmic reticulum is the core, and specific tethered proteins at the membrane contact site bind to the organelle membrane and various protein complexes work together to perform specific functions, such as lipid transport, Ca2+ transfer, etc. This review studies on the structure and function of membrane contact sites and their key roles in organelle interactions, focusing on the connection between the endoplasmic reticulum and plasma membrane, mitochondria and Golgi, as well as the association between the key proteins at membrane contact sites and the occurrence and development of various diseases.

Efficacy and safety of anticoagulant therapy with warfarin in hemodialysis patients with atrial fibrillation / 中华全科医师杂志

Objective To evaluate the efficacy and safety of anticoagulant therapy with warfarin for hemodialysis patients with atrial fibrillation.Methods A cohort of 118 hemodialysis patients with atrial fibrillatio were enrolled.The patients were divided into two groups:in warfarin group (n=53) the standard medication of warfarin was given for at least 3 months by adjusting the INR between 2.0 and 3.0;while no anticoagulants were given after discharge in non-warfarin group (n=65).Patients were followed up regularly,the events of stoke,death and bleeding were documented and compared between two groups.Results There were no significant differences in age,sex,underlying disease,predictive score of stroke risk in patients with non-valvular atrial fibrillation (CHA2DS2-VASc),predictive score of bleeding risk in patients with non-valvular atrial fibrillation (HAS-BLED) and dialysis frequency between the two groups (P＞0.05).There were no significant differences in events of ischemic stroke [11％(6/53) vs.12％(8/65),x2=0.027,P=0.87],bleedings [(9％(5/53) vs.5％(3/65),P=0.46] and gastrointestinal bleeding events [6％(3/53) vs.3％ (2/65),P=0.66] between two groups.There were no significant differences in the death event [15％(10/53) vs.22％(14/65),x2=0.129,P=0.72] and cardiac death [9％(5/53) vs.11％(7/65),x2=0.057,P=0.81] between two groups.Conclusion This study suggests that warfarin may not prevent ischemic stroke in hemodialysis patients with chronic atrial fibrillation,further studies are needed to determine the risks and benefits of anticoagulation therapy in these patients.

Effects of schisandrin B on pharmacokinetics of doxorubicin in rats / 中国药科大学学报

@#To investigate the effect of combination of schisandrin B and doxorubicin on the pharmacokinetic behavior of doxorubicin in SD rats. An LC-MS/MS method was established for the determination of doxorubicin and its main metabolite doxorubicinol in SD rats plasma. Separation was performed on Agilent Eclipse XDB-C18 column(100 mm×2. 1 mm, 3. 5 μm)using 0. 1% formic acid solution and acetonitrile as mobile phase with a liner gradient program. The ion transitions were performed under ESI position model at m/z 544. 2→397. 3(doxorubicin), m/z 546. 2→399. 2(doxorubicinol), m/z 528. 2→381. 2(daunorubicin, internal standard). Calibration curves of doxorubicin(0. 500-500 ng/mL)and doxorubicinol(0. 200-50. 0 ng/mL)showed good linear regression. The precision and accuracy met the requirements. The variation coefficient of extraction recovery and matrix effect was less than 15%. The AUC0-t of doxorubicin were(605. 69±145. 84)and(564. 53±23. 99)ng ·h/mL in alone and combined group, respectively. The AUC0-t of doxorubicinol were(26. 69±13. 41)and(29. 00±2. 78)ng ·h/mL, respectively. Results indicated that, schisandrin B had not affected the pharmacokinetic behavior of doxorubicin in SD rats.

Research progress of P-glycoprotein induction / 中国药科大学学报

P-glycoprotein(P-gp)is an important efflux protein of ATP-binding cassette transporter superfamily. Cells could be protected from detrimental xenobiotics by the up-regulation of efflux pumps.In this review, an extensive literature search for P-gp induction research was conducted, and a focus was brought onto the P-gp induction models,experiment methods and its applications in drug discovery.We mainly introduced the in vitro cell-based models and in vivo rodent animal models for induction research, methods that investigate induction potency by detecting the protein,gene expression and efflux function,as well as co-regulation between P-gp and other transporters or drug metabolism enzymes.P-gp induction can serve as a clinical therapeutic strategy by reducing the intracellular concentration of deleterious xenobiotics significantly,and the in silico P-gp induction pharmacophore model was also discussed.This review could be of great importance for pre-clinical drug design, the screening of new synthesized compounds and the prediction of potential clinical drug-drug interactions.

Mid-term effects of arthroscopic anatomical reconstruction of double bundles of the anterior cruciate ligament / 中华创伤骨科杂志

Objective To investigate the mid-term clinical outcomes of arthroscopic anatomical re-construction of double bundles of the anterior cruciate ligament ( ACL ) . Methods The clinical data of 78 patients diagnosed with ACL rupture from April 2012 to July 2014 were analyzed retrospectively. They were 60 males and 18 females, aged from 19 to 56 years ( mean, 26. 8 years ) . The time from injury to surgery ranged from one week to 23 months ( mean, 5. 8 months ) . All of them obtained positive results in anterior drawer test and Lachman test preoperatively. Their preoperative KT-1000 examinations showed an average side-to-side difference of 8. 29 ± 1. 81 mm in anterior laxity. They were all treated with arthroscopic anatomical recon-struction of double bundles of the anterior cruciate ligament using autologous hamstrings. The International Knee Documentation Committee ( IKDC ) and Lysholm scores were used to assess their knee function at the last follow-up. Results All the 78 patients were followed up for 34. 6 months on average ( range, from 25 to 56 months ) . At the last follow-up, the IKDC and Lysholm scores were significantly increased from preoper-ative 42. 6 ± 9. 5 and 44. 4 ± 8. 5 to postoperative 92. 9 ± 2. 8 and 94. 2 ± 3. 4, respectively ( P <0. 05 ) . The Lachman test was negative in 73 cases ( 93. 6％) and the pivot shift test was negative in 69 cases ( 88. 5％) . The KT-1000 examinations showed that the side-to-side difference in anterior laxity averaged 1. 47 ± 0. 68 mm, significantly improved from the preoperative values ( P <0. 05 ) . At their last follow-up, 29 patients underwent MRI scans which showed continuity of the anteromedial and posterolateral bundles of the anterior cruciate liga-ment. Conclusion The arthroscopic anatomical reconstruction of double bundles of the anterior cruciate ligament can restore the knee stability and achieve fine mid-term clinical outcomes.

Identification of major constituents from propolis by LC-IT-TOF-MS / 中国药科大学学报

A qualitative analytical method of liquid chromatography coupled with ion trap time-of-flight mass spectrometry (LC-IT-TOF/MS) was developed for identification of major constituents in propolis from China (Shandong).The LC-IT-TOF-MS was performed on a Shim-pack VP-ODS column (2.0 mm× 150 mm,5 μm) with the mobile phase consisting of acetonitrile and water containing 0.2％ formic acid in gradient mode,and the flow rate was set at 0.3 mL/min.Negative ion mode was used for IT-TOF-MS.According to the accurate molecular weight,MS fragment pathway,comparison with the retention time of reference compounds,total 31 compounds,including twenty-five phenolic acids and six flavonoids were identified.The LC-IT-TOF-MS qualitative analysis method can be used for analyzing major components of propolis quickly and accurately.

Rhein:a novel matrix for profiling and imaging of endogenous metabolites by matrix-assisted laser desorption/ionization-mass spectrometry / 中国药科大学学报

@#MALDI-MS is an emerging technique that enables rapid profiling of endogenous substances in a high throughput. Nevertheless, its application to metabolite analysis is often hindered by the presence of massive matrix background peaks using conventional matrices. Herein rhein was introduced as a novel matrix for MALDI-MS analysis of cation metabolites. Several strong and weak basic metabolites were measured using rhein. Compared to the previously reported ionless matrix NSA and conventional matrix CHCA, clean MALDI-MS spectra were obtained devoid of matrix signals. Subsequently, we applied this matrix to profile a biologically complex sample, mice intestinal contents. In addition, we have validated the applicability of rhein to MALDI-MS imaging, which allows for simultaneous mapping of hundreds of metabolites from single mouse ileum section. In summary, the discovery of rhein as a novel MALDI matrix enables efficient and reliable profiling and imaging of endogenous metabolites from biological samples with no matrix interferences, which will greatly promote the utility of MALDI-MS-based platform for metabolome studies.

Shenmai injection inhibits infIammatory response in IipopoIysaccharides-in-duced septic mice / 中国药科大学学报

This study was to investigate the regulation of lipopolysaccharides(LPS)-induced sepsis in mice by preadministration of Shenmai injection (SMI)and the therapeutic differences between male and female.The females and males were randomly grouped by weight,including control group,LPS-induced sepsis model group and SMI administration group.After preadministration of SMI for 14 days,10 mg/kg LPS were intraperitoneally injected subsequently to induce sepsis.The survival rate of mice,level of serum cytokines and the mRNA expres-sion of proinflammatory cytokines in main tissues were detected to evaluate the impact of SMI in LPS-induced sepsis mice.From the survival rate,which is considered as a gold standard of improvement in sepsis,significant protective effect can be observed after SMI pretreatment in LPS-induced sepsis mice,a more significant effect was shown in the females.Consisting with the serum cytokines levels,SMI significantly inhibited proinflammatory cyto-kines including IL-6,IL-1βand TNF-αmRNA expression in tissues and the regulation of IL-6 was most signifi-cant,which was consistent with the results of ELISA in serum.Moreover,the liver tissue acquired a more evident impact than any other tissues,which fits with the ratio of dry/wet weight.SMI can significantly inhibit inflammato-ry response by delivery in advance in LPS-induced septic mice,providing strong evidence for elaborating mecha-nism in the treatment of cardiovascular disease related inflammation and shock.