ObjectiveTo identify the prototypical components and metabolites absorbed into blood and cerebrospinal fluid of Schisandrae Chinensis Fructus（SCF） based on sequential metabolism combined with liquid chromatography-mass spectrometry. MethodBlood and cerebrospinal fluid samples of integrated metabolism， intestinal metabolism and hepatic metabolism were collected from male SD rats after gavage and in situ intestinal perfusion administration， and ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry（UPLC Q-Exactive Orbitrap MS） was used to analyze and compare the differences in the spectra of SCF extract， blank plasma， administered plasma， blank cerebrospinal fluid and administered cerebrospinal fluid with ACQUITY UPLC BEH Shield RP18 column（2.1 mm×100 mm， 1.7 µm）， the mobile phase was acetonitrile（A）-0.1% formic acid aqueous solution（B） for gradient elution（0-7 min， 95%B； 7-12 min， 95%-35%B； 12-17 min， 35%-15%B； 17-20 min， 15%-12%B； 20-22 min， 12%-5%B； 22-23 min， 5%B； 23-25 min， 5%-95%B； 25-28 min， 95%B）. And heated electrospray ionization（HESI） was used with positive and negative ion modes， the scanning range was m/z 100-1 500. The prototypical constituents and their metabolites absorbed into blood and cerebrospinal fluid of SCF were identified according to the retention time， characteristic fragments， molecular formulae and the information of reference substances. ResultA total of 42 chemical components were identified in the extract of SCF， including lignans， flavonoids， amino acids， tannins， and others， of which lignans were the main ones. A total of 27 prototypical components and 14 metabolites were identified in plasma samples from different sites. A total of 15 prototypical components and 9 metabolites were identified in cerebrospinal fluid. The main metabolic reactions involved in the formation of metabolites were mainly demethylation， methylation， demethoxylation and hydroxylation. ConclusionThrough the systematic identification of the prototypical components and metabolites of SCF in rats， it provides data support for further better exploring the material basis of SCF in the treatment of central nervous system diseases.

Objective:To evaluate direct and indirect costs for schizophrenia,major depressive disorder(MDD)and bipolar disorder,and to compare their differences of cost composition,and to explore the drivers of the total costs.Methods:A total of 3 175 inpatients with schizophrenia,MDD,and bipolar disorder were recruited.In-patient's self-report total direct of medical costs outpatient and inpatient,out-of-pocket costs,and direct non-medical costs were regarded as direct costs.Productivity loss and other loss caused by damaging properties were defined as indirect costs.The perspectives of this study included individual and societal levels.Multivariate regression analysis was applied for detecting the factors influencing disease costs.Results:The total cost of schizophrenia was higher than those of MDD and bipolar disorder at individual and societal levels.The indirect costs of three mental disorders were higher than the direct costs,and the indirect cost ratio of bipolar disorder was higher than those of schizophre-nia and MDD.Age,gender,working condition and marital status(P＜0.05)were the important drivers of total costs.Conclusion:The economic burden of the three mental disorders is relatively heavy.Schizophrenia has heaviest disease burden,and the productivity loss due to mental disorders is the driving force of the soaring disease cost

Objective:To compare demographic characteristics,clinical characteristics,therapeutic characteris-tics and physiological indicators of patients with bipolar Ⅰ disorder and bipolar Ⅱ disorder.Methods:A total of 381 patients with bipolar disorder(BD)diagnosed by the Diagnostic and Statistical Manual of Mental Disorders 5 th Edi-tion(DSM-5)were selected,including 302 patients with BD-Ⅰ(79.27％),74 patients with BD-Ⅱ(19.42％)and 5 patients with other specific and related disorders(1.31％).Demographic and clinical characteristics were collected with self-designed clinical information questionnaire.Multivariate logistic regression and multivariate linear regres-sion analysis were used for analysis.Results:Compared with patients with BD-Ⅱ,patients with BD-Ⅰ had more risk to have psychotic features(OR=5.75,95％CI:2.82-11.76),longer disease duration,and more repeated transcra-nial magnetic therapy(OR=3.09,95％CI:1.02-9.35),higher uric acid,total cholesterol and high-density lipo-protein.BD-Ⅰ in Han nationality was more common(OR=11.50,95％CI:1.76-75.30),and had lower education level(OR=10.22,95％CI:1.16-89.77),and less family history of psychosis(OR=2.34,95％CI:1.01-5.42).Conclusion:There are significant differences between BD-Ⅰ and BD-Ⅱ in demographic and clinical charac-teristics,treatment status,and physiological indicators,which could provide clues for exploring the pathogenesis of bipolar disorder.

Objective:To investigate the mechanism of Porphyridium cruentum polysaccharides extract in reducing telangiectasis.Methods:Experiments were conducted between January 10, 2023, and April 18, 2023, at the Yunnan Characteristic Plant Extraction Laboratory. In vitro experiments were conducted to investigate the effect of different concentrations of Porphyridium cruentum polysaccharides extract (Porphyridium cruentum Pro) on tube formation and migration of microvascular endothelial cells, using tube formation assay and migration assay. In an in vivo experiment, a chick embryo chorioallantoic membrane assay was conducted to assess the impact of Porphyridium cruentum Pro on vasoconstriction. Finally, a western blot analysis was conducted to confirm the impact of Porphyridium cruentum Pro on increasing the protein level of endothelin-1 in microvascular endothelial cells.Results:When the concentration of Porphyridium cruentum Pro was below 1.0%, it did not significantly affect all the stages of tube formation and the migration ability of microvascular endothelial cells ( P>0.05). However, at a concentration of 3.0%, there was a noticeable effect on the collapse of the formed tubes. In comparison to the control group, some tubes started collapsing 24 hours post-treatment, despite the lack of statistical significance in the analysis. After 48 hours of treatment, all the tubes collapsed, as evidenced by a significant decrease in the number of tubes ( t=9.02, P<0.05) and total tube length ( t=10.89, P<0.05) per field. The migration assay study revealed that at a concentration of 3%, there was observed cell detachment and atrophy, while the cell migration capacity of microvascular endothelial cells remained relatively unchanged. When treated with a 3.0% concentration of Porphyridium cruentum Pro for 30 minutes, a noteble vasoconstriction was observed in the blood vessels of the chick embryo chorioallantoic membrane. Moreover, after a 24-hour treatment with 0.3% Porphyridium cruentum Pro, there was an elevation in endothelin-1 protein expression level in microvascular endothelial cells. Conclusions:The observed function of Porphyridium cruentum Pro in reducing telangiectasis may associated to its ability to enhance the expression of endothelin-1 in vascular endothelial cells, which in turn promotes the vasoconstriction. Meanwhile, the angiogenesis is not significantly affected by Porphyridium cruentum Pro.

Objective:To simulate the occupancy rates of baicalein， quercetin and galangin on the target sites of xanthine oxidase in vivo. Method:In this experiment， the half inhibitory concentration （IC₅₀） of febuxostat， baicalein， quercetin and galangin against xanthine oxidase were determined by in vitro enzymatic reaction. Binding free energy was predicted by molecular docking technology and their association rate constant （k_on） and dissociation rate constant （k_off） were determined by surface plasmon resonance technology. Based on measured binding kinetic parameters （k_on and k_off） and extracted pharmacokinetic data， the target occupancy model in vivo was established. Result:The IC₅₀ values of febuxostat， baicalein， quercetin and galangin were 0.002 7， 1.63， 0.38， 1.59 µmol·L^-1， respectively. The IC₅₀ of febuxostat was very close to that reported in the literature. The predicted curve of target occupancy rate in vivo of febuxostat was consistent with its duration of clinical efficacy. When single intragastric administration of long-circulating liposomes of quercetin with dose of 100 mg·kg^-1 in rats， the time of target occupancy rate >70% in vivo lasted for about 3.9 h. When rats were orally administered baicalein and galangin with dose of 200 mg·kg^-1， the time of target occupancy rate >50% in vivo lasted for about 10 h and 1.7 h， respectively. Conclusion:The prediction model of xanthine oxidase target occupancy constructed by drug target binding kinetics and in vivo pharmacokinetic curves can effectively evaluate the in vivo inhibitory activity of compounds against the target.

Based on the target occupancy mathematical model, the binding kinetic process of potential active ingredients of lowering uric acid in Chrysanthemum morifolium with xanthine oxidase(XOD) was evaluated. The potential active ingredients of lowering uric acid in Ch. morifolium were screened by UPLC-Q-Exactivems MS technology, reference substance identification and in vitro enzymatic kinetics experiments. The binding kinetic parameters of xanthine oxidase and potential inhibitor in Ch. morifolium were determined by surface plasma resonance(SPR). The verified mathematical model of the XOD target occupancy evaluated the kinetic binding process of inhibitors and xanthine oxidase in vivo. According to UPLC-Q-Exactive MS and reference substance identification, 39 potential uric acid-lowering active ingredients in Ch. morifolium extracts were identified and the inhibitory activities of 23 compounds were determined. Three potential xanthine oxidase inhibitors were screened, namely genistein, luteolin, and apigenin. whose IC_(50 )were 1.23, 1.47 and 1.59 μmol·L~(-1), respectively. And the binding rate constants(K_(on)) were 1.26×10~6, 5.23×10~5 and 6.36×10~5 mol·L~(-1)·s~(-1), respectively. The dissociation rate constants(K_(off)) were 10.93×10~(-2), 1.59×10~(-2), and 5.3×10~(-2 )s~(-1), respectively. After evaluation by different administration methods, the three selected compounds can perform rapid and sustained inhibition of xanthine oxidase in vivo under combined administration. This study comprehensively evaluated the target occupancy process of three effective components in different ways of administration in vivo by UPLC-MS, concentration-response method, SPR technology and xanthine oxidase target occupancy model, which would provide a new research idea and method for screening active ingredients in traditional Chinese medicine.
Chromatography, Liquid ; Chrysanthemum ; Flavonoids ; Kinetics ; Pharmaceutical Preparations ; Tandem Mass Spectrometry ; Xanthine Oxidase/metabolism*

OBJECTIVE： To screen the quality control components of Chrysanthemum morifolium based multiple component metabolism， and study its network pharmacology effect. METHODS： The water extract of C. morifolium was prepared. A total of one rats were selected， water extract of C. morifolium was perfused in jejunum segment after abdominal anesthesia； plasma sample 1 was collected by double perfusion collection. Other 3 rats were given water extract of C. morifolium intragastrically， and plasma sample 2 was collected by abdominal aorta blood collection. UPLC-MS/MS was used to analyze water extract of C. morifolium and plasma sample component， and prototype blood-entry component in water extract of C. morifolium was identified after metabolism. TCMSP and Swiss Target Prediction database were used to screen the core target of prototype blood-entry component. DAVID database was used to enrich the related pathways of core target. The quality control components were screened according to topological parameters. Cytoscape software was used to analyze pharmacological effect of quality control components of C. morifolium. RESULTS： After UPLC-MS/MS analysis， 27 compounds were identified in water extract of C. morifolium， among which there were 12 prototype blood-entry components. After network pharmacology analysis， 7 quality control components were identified， i.e. cosmosiin， apigenin-7-O-glucuronide， luteolin， tilianin， apigenin， hesperetin， acacetin. It was possible to treat cancer， cardiovascular and cerebrovascular diseases， and neurological diseases by acting on metabolic pathway， cancer related pathway， signal transduction related pathway， adipocyte lipolysis regulatory pathway， etc. CONCLUSIONS： The study screen the possible quality control components of water extract of C. morifolium. The theoretical pharmacological effect of it can be clarified through network pharmacology， which can provide a new idea for the utilization of C. morifolium.

Objective To study an improved isolated method of single human atrial myocytes.Methods Enzyme digestion method was used to isolate single myocytes from human atrial and whole-cell patch clamp technique was used to record small conductance calcium activated potassium current.Results This method obtained a large number of atrial myocytes.The total amount of atrial myocytes in SR group was 320±30 while AF group was 230±20 and the difference was statistically significant(P<0.01).In this study,a large number of simple and striated single atrial myocytes were obtained,and a typical small-conductance calcium-activated potassium channel current was recorded on the isolated atrial myocytes.Conclusion The established isolated method is simple,stable and effective.We can acquire a large amount of single atrial myocytes with good quality.

Aim To investigate if LPS increases the sterol regulatory element binding proteins ( SREBPs ) cleavage-activating protein ( SCAP )-SREBP2 expres-sion by activation of mTOR signal pathway in THP-1 macrophages , upgrading LDLr level , causing foam-cell formation .Methods THP-1 macrophages were incu-bated in serum free medium in the absence of 5 mg?L-1 LDL alone , or 5 mg? L-1 LDL plus 200 μg? L-1 LPS, or 5 mg? L-1 LDL plus 200 μg? L-1 LPS plus 10 μg? L-1 rapamycin .Morphological examination of macrophages was performed with Oil Red O staining . Expression changes of LDLr , SREBP2, SCAP, S6K1 and mTOR mRNA were detected by real time quantita-tive polymerase chain reaction ( PCR ) .Western blot was used to analyze protein expression changes of LD-Lr, S6K1 and mTOR.Translocation of SCAP-SREBP2 complex from the endoplasmic reticulum ( ER ) to the Golgi was determined by confocal microscopy .Results LPS enhanced transformation of THP-1 macrophages into foam cells by increased uptake of lipid as evi-denced by Oil Red O assay .LPS increased mRNA lev-els of LDLr, SREBP2, SCAP, S6K1 and mTOR ( P <0.05) .Rapamycin reduced the mRNA levels of LDLr , SREBP2,SCAP,S6K1 and mTOR induced by LPS ( P<0.05 ) .Western blot demonstrated that LPS also caused over-expression of protein of LDLr , S6K1 and mTOR(P<0.05).Rapamycin reduced the expression of protein of LDLr, S6K1 and mTOR induced by LPS ( P <0.05 ) .Confocal microscopy demonstrated LPS caused an escape of SCAP-SREBP2 complex from the ER to the Golgi .Rapamycin inhibited the translocation of SCAP-SREBP2 complex from the ER to the Golgi . Conclusions Inflammatory stress increases SCAP/SREBP2 expression by activation of mTOR signal path-way, resulting in an escape of SCAP-SREBP2 complex from the ER to the Golgi , furthermore elevating LDLr expression and causing foam-cell formation .Rapamy-cin reverses the activation of mTOR signal pathway and decreases lipid deposition in THP-1 macrophages in-duced by LPS .

G protein-coupled bile acid receptor 1(TGR5) is a specific membrane receptor of bile acids , playing an important role in the bile acid signaling network .Its activation has been proved to increase the glycemic control , regulate of blood lipid balance , enhance energy expenditure , exert anti-inflammatory actions and so on .It suggests that TGR5 may play an important role in metabolic diseases .