Effects of different light sources on the stability of cinepazide maleate in sodium chloride solution / 中国基层医药

Objective:To investigate the effects of different light sources on the stability of cinepazide maleate in sodium chloride solution, providing a theoretical basis for standardizing operation for avoiding light in clinical medication.Methods:First, a standard curve was created and methodological studies were conducted. Then, intravenous drip solutions were prepared using clinically prescribed methods. Then, the absorbance of the solution of cinepazide maleate injection under outdoor light, light emitting diode (LED) light, indoor light, artificial sunlight with a color temperature of 6 500 K (D65), and under a shading condition for 1, 2, 3, and 7 hours were brought into the standard curve to obtain the concentrations, and the change of the concentrations was studied.Results:There was no light decomposition phenomenon of cinepazide maleate injection in the shading group (control group) within 7 hours. In the other four experimental groups, there was no obvious light decomposition phenomenon of cinepazide maleate injection in the LED light group. Compared with the shading group, cinepazide maleate injection in the indoor, outdoor, and D65 groups began to exhibit a light decomposition phenomenon at 1 hour ( F = 44 840.44, P < 0.001). At 2 hours, cinepazide maleate content in the outdoor and D65 groups began to decrease significantly compared with the shading group ( F = 15 459.12, P < 0.001). At 7 hours, cinepazide maleate exhibited significantly greater light decomposition in the outdoor group [(29.84 ± 0.43) L·mol -1·cm -1] and D65 group [(21.01 ± 0.51) L·mol -1·cm -1] than the shading group [(101.65 ± 1.5) L·mol -1·cm -1] ( F = 63 106.32, P < 0.001). There was no significant difference in cinepazide maleate content between LED and shading groups ( P > 0.05). Conclusion:Cinepazide maleate injection is stable under the common LED light source and therefore it is not necessary to take lightproof operation. Cinepazide maleate injection is unstable under indoor, outdoor, and D65 light sources, and it is necessary to take lightproof operation.

Compatibility stability of cinepazide maleate injection and dopamine hydrochloride injection / 中国基层医药

Objective:To investigate the compatibility stability of cinepazide maleate injection and dopamine hydrochloride injection and explore the basis for the combined application of cinnamazide maleate injection and dopamine hydrochloride injection.Methods:A method for determining cinnamazide maleate injection and dopamine hydrochloride injection was established using an ultraviolet-visible spectrophotometer and verified from March 1, 2021 to May 20, 2021. The color and pH value of the solution prepared using the two drugs were determined within 5 hours at room temperature. The content change of the prepared solution was determined using an ultraviolet-visible spectrophotometer.Results:The linear range of the mass concentration of cinnamazide maleate was 4.03 - 32.24 mg/L and the linear range of dopamine hydrochloride was 20 -120 mg/L. At 25 ℃, the color of the prepared solution did not change within 5 hours and the pH value was in the range of 4.43 ± 0.06, indicating that the pH of the prepared solution did not change markedly. The concentrations of cinnamazide maleate and dopamine hydrochloride were (98.23 ± 1.09)% and (99.96 ± 0.41)% respectively, indicating good stability.Conclusion:The prepared solution using cinepazide maleate injection and dopamine hydrochloride injection can be used within 5 hours at 25 ℃.

Value and research progress of BRAFV600E mutation in the diagnosis and treatment of thyroid carcinoma / 国际外科学杂志

Thyroid Carcinoma, as one of the most common malignant tumors in the endocrine system and head and neck, has a rising incidence rate in the world in recent years, which seriously affects human health. Thyroid carcinoma is often divided into 4 pathological types: papillary carcinoma, follicular carcinoma, medullary carcinoma and undifferentiated carcinoma. Among them, papillary carcinoma is the most common with low malignancy and undifferentiated carcinoma is extremely rare with the highest malignancy. BRAFV600E mutation has been found to be closely related to the genesis, development mechanism and prognosis of thyroid carcinoma, and is a current molecular research focus. Clinically, BRAFV600E mutation is often considered as a molecular marker affecting the prognosis of thyroid carcinoma. In this paper, the relevant literature in recent years was reviewed, mainly from the aspects of BRAFV600E mutation and thyroid carcinoma diagnosis, clinicopathological features, guidance of clinical treatment decision, prognosis and so on, to evaluate the clinical application value of BRAFV600E mutation.

Simultaneous determination of eleven volatile components in Cinnamomi Oleum by GC-MS / 中国中药杂志

A gas chromatography-triple quadrupole mass spectrometry(GC-MS) method was established for the simultaneous determination of eleven volatile components in Cinnamomi Oleum and the chemical pattern recognition was utilized to evaluate the quality of essential oil obtained from Cinnamomi Fructus medicinal materials in various habitats. The Cinnamomi Fructus medicinal materials were treated by water distillation, analyzed using GC-MS, and detected by selective ion monitoring(SIM), and the internal standards were used for quantification. The content results of Cinnamomi Oleum from various batches were analyzed by hierarchical clustering analysis(HCA), principal component analysis(PCA), and orthogonal partial least squares-discriminant analysis(OPLS-DA) for the statistic analysis. Eleven components showed good linear relationships within their respective concentration ranges(R~2>0.999 7), with average recoveries of 92.41%-102.1% and RSD of 1.2%-3.2%(n=6). The samples were classified into three categories by HCA and PCA, and 2-nonanone was screened as a marker of variability between batches in combination with OPLS-DA. This method is specific, sensitive, simple, and accurate, and the screened components can be utilized as a basis for the quality control of Cinnamomi Oleum.

Content determination of seven active components of Eucommiae Cortex in aortic vascular endothelial cells of spontaneously hypertensive rats by UPLC-MS/MS / 中国中药杂志

In the present study, the contents of seven active components [genipinic acid(GA), protocatechuic acid(PCA), neochlorogenic acid(NCA), chlorogenic acid(CA), cryptochlorogenic acid(CCA),(+)-pinoresinol di-O-β-D-glucopyranosid(PDG), and(+)-pinoresinol 4'-O-β-D-glucopyranoside(PG)] of Eucommiae Cortex in aortic vascular endothelial cells of spontaneously hypertensive rats(SHR) were simultaneously determined by ultra-high liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS). The qualified SHR models were selected. The primary aortic endothelial cells(VECs) of rats were separated and cultured by ligation and adherence, followed by subculture. After successful identification, an UPLC-MS/MS method for simultaneously determining the contents of GA, PCA, NCA, CA, CCA, PDG, PG in seven components of Eucommiae Cortex in VECs was established, including specificity, linearity, matrix effect, recovery, accuracy, precision and stability. The established method had the lo-west limit of quantification of 0.97-4.95 μg·L~(-1), accuracy of 87.26%-109.6%, extraction recovery of 89.23%-105.3%, matrix effect of 85.86%-106.2%, and stability of 86.00%-112.5%. Therefore, the established accurate UPLC-MS/MS method could rapidly and simultaneously determine the contents of the seven active components of Eucommiae Cortex in VECs of SHRs, which provided a refe-rence for the study of cellular pharmacokinetics of active components of Eucommiae Cortex extract.

The two-stage interaction of Ebola virus VP40 with nucleoprotein results in a switch from viral RNA synthesis to virion assembly/budding

Ebola virus (EBOV) is an enveloped negative-sense RNA virus and a member of the filovirus family. Nucleoprotein (NP) expression alone leads to the formation of inclusion bodies (IBs), which are critical for viral RNA synthesis. The matrix protein, VP40, not only plays a critical role in virus assembly/budding, but also can regulate transcription and replication of the viral genome. However, the molecular mechanism by which VP40 regulates viral RNA synthesis and virion assembly/budding is unknown. Here, we show that within IBs the N-terminus of NP recruits VP40 and is required for VLP-containing NP release. Furthermore, we find four point mutations (L692A, P697A, P698A and W699A) within the C-terminal hydrophobic core of NP result in a stronger VP40-NP interaction within IBs, sequestering VP40 within IBs, reducing VP40-VLP egress, abolishing the incorporation of NC-like structures into VP40-VLP, and inhibiting viral RNA synthesis, suggesting that the interaction of N-terminus of NP with VP40 induces a conformational change in the C-terminus of NP. Consequently, the C-terminal hydrophobic core of NP is exposed and binds VP40, thereby inhibiting RNA synthesis and initiating virion assembly/budding.

Simultaneous determination of content of eight components in Caesalpinia decapetala by UPLC-MS/MS / 中国中药杂志

The present study established the ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for simultaneous determination of the content of eight major active components in Caesalpinia decapetala and performed the quality evaluation of C. decapetala from different habitats with the chemical pattern recognition. The analysis was carried out on a Waters BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) at 40 ℃, with the mobile phase of water containing 0.1% formic acid(A) and acetonitrile containing 0.1% formic acid under gradient elution, the flow rate of 0.3 mL·min~(-1), and the injection volume of 1 μL. The electrospray ionization(ESI) source in the negative mode and multiple reaction monitoring(MRM) were used for MS quantitative analysis. The content results were analyzed by the hierarchical cluster analysis(HCA) and principal component analysis(PCA) for the evaluation of the quality difference. Eight components showed good linear relationships within their respective concentration ranges(r>0.999), with the average recoveries of 96.85%-103.4% and RSD of 0.52%-2.8%. The analysis results showed that the quality of samples from different batches was different. The samples were classified into three clusters by HCA and PCA. The method is simple, sensitive, accurate, and efficient, and can be used for the quality evaluation of C. decapetala.

Difference Analysis of Pharmacokinetic Behavior of Active Ingredients in Wujiwan Between Normal Rats and Rats with Irritable Bowel Syndrome / 中国实验方剂学杂志

Objective:To establish an ultra-performance liquid chromatography-mass spectrometry （UPLC-MS/MS） for determining the plasma concentrations of 10 active ingredients in Wujiwan at different time points after oral administration， and to compare the pharmacokinetic characteristics between normal rats and rats with chronic visceral hypersensitive irritable bowel syndrome （CVH-IBS）. Method:CVH-IBS rat model was prepared by the neonatal rat colon percutaneous transluminal coronary angioplasty （PTCA） balloon stimulation method. After intragastric administration of Wujiwan （0.245 g·kg^-1）， blood was collected from the jugular vein at different time points， and the plasma concentrations of 10 active ingredients （berberine hydrochloride， palmatine hydrochloride， coptisine hydrochloride， jatrorrhizine hydrochloride， epiberberine， dihydroberberine， evodiamine， evodine， paeoniflorin， albiflorin） in Wujiwan was detected simultaneously by UPLC-MS/MS， the pharmacokinetic parameters of each component in normal rats and CVH-IBS rats were calculated. Result:The established UPLC-MS/MS could sensitively and accurately detect the plasma concentrations of 10 active ingredients of Wujiwan in rats. Compared with the normal group， the absorption rates of these 10 active ingredients of Wujiwan in the blood of CVH-IBS rats all decreased to a certain extent， and the peak time （<italic>t</italic>_max） was prolonged. Among them， the <italic>t</italic>_max of berberine hydrochloride and jatrorrhizine hydrochloride were significantly prolonged from 54 minute and 39 minute to 90 minute， respectively （<italic>P</italic><0.05， <italic>P</italic><0.01）. Area under the plasma concentration-time curve （AUC_0-<italic>_t</italic>） of each component increased， and evodiamine and paeoniflorin were significantly different （<italic>P</italic><0.05，<italic> P</italic><0.01）. The clearance rates （CL/<italic>F</italic>） of these 10 active ingredients were all decreased， among which berberine hydrochloride， palmatine hydrochloride and evodiamine had significant differences （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:There are significant differences in the pharmacokinetic behavior of the active ingredients in Wujiwan between normal rats and CVH-IBS rats， which may be related to the destruction of microstructure of intestinal epithelial cells and the change of activity of liver enzymes under the pathological state of IBS.

Analysis of research status based on projects in pharmacokinetics of traditional Chinese medicine funded by National Natural Science Foundation of China / 中国中药杂志

The pharmacokinetics of traditional Chinese medicine is a subject that studies the dynamic changes of the absorption, distribution, metabolism and excretion of complex components of traditional Chinese medicine, which is an important method for elucidating the pharmacodynamic material basis, action characteristics, and compatibility mechanisms of traditional Chinese medicine. However, given on the fact that traditional Chinese medicine is a multi-dimensional and complex system with multiple components, multiple pathways, multiple targets, and an unclear pharmacodynamic material basis, the research on the pharmacokinetics of traditio-nal Chinese medicine has become a scientific and technological problem. Although the pharmacokinetics of traditional Chinese medicine has achieved remarkable development with the emergence of new theories, methods and technologies, there are still some problems in the application of the research direction of the pharmacokinetics of traditional Chinese medicine judging from the current application of the National Natural Science Foundation of China. Therefore, this article discussed the current research status on pharmacokinetics of traditional Chinese medicines by analyzing the projects funded by the National Natural Science Foundation of China in the past 5 years from 2016 to 2020, mainly including the application and funding analysis, main research contents of the projects in pharmacokinetics of traditional Chinese medicines. And the research hotspots, difficulties and deficiencies were focused in order to provide certain reference for researchers engaged in pharmacokinetics of traditional Chinese medicine.

Controversy and progress on whether to retain left colonic artery in radical resection of rectal cancer / 中华胃肠外科杂志

Japanese Society for Cancer of the Colon and Rectum (JSCCR) guideline 2019 recommended that lymph node dissection for advanced rectal cancer should include the lymphatic adipose tissue at the root of the inferior mesenteric vessels, but the ligation site of the inferior mesenteric artery (IMA) was not determined, and the NCCN guideline did not indicate clearly whether to retain the left colonic artery (LCA). Controversy over whether to retain LCA is no more than whether it can reduce the incidence of anastomotic complications or postoperative functional damage without affecting the patients' oncological outcome. Focusing on the above problems, this paper reviews the latest research progress. In conclusion, it is believed that the advantages of retaining LCA are supported by most studies, which can improve the blood supply of the proximal anastomosis, and technically can achieve the same range of lymph node dissection as IMA high ligation. However, whether it affects the survival of patients, reduces the incidence of anastomotic leakage, and improves the quality of life of patients, more high-quality evidence-based medical evidence is still needed.

Simultaneous determination of four components of Shuganning Injection in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study / 中国中药杂志

This study aims to reveal the pharmacokinetics of Shuganning Injection in normal rats. In this experiment,ultra-high performance liquid chromatography-electrospray-tandem mass spectrometry( UPLC-ESI-MS/MS) was used to establish an analytical method for simultaneous determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma. Then,the non-compartmental model( NCA) in Phoenix WinN onL in 6. 4 software was used to fit pharmacokinetic parameters. The methodological validation showed that the linear relationship of the components in rat plasma samples were good( r>0. 995). The recovery rate and matrix effect of plasma samples with low,middle and high concentration were 79. 14%-101. 4%. The intra-day and inter-day precision,accuracy and stability meet the requirements of biological sample analysis. The half-life( t1/2) of chlorogenic acid,gardenioside,oroxylin A did not change significantly and the area under blood concentration-time curve( AUC0-t) is proportional to the dose,which suggested that three components showed a linear kinetic characteristics,but baicalin showed nonlinear kinetic characteristics. Moreover,the retention time of each component in rats was short. The established UPLC-MS/MS quantitative analysis method is rapid,sensitive and accurate,which can be used for the determination of chlorogenic acid,gardenioside,oroxylin A and baicalin in rat plasma and pharmacokinetic study of Shuganning Injection.

Study on metabolites of Laportea bulbifera extract in rat feces based on UHPLC-Q-TOF-MS~E technique / 中国中药杂志

This project is to study the metabolites of Laportea bulbifera extract in rat feces. After the SD rats were gavaged with the extract(136 g·kg~(-1), according to the crude drug dose), the metabolites in their feces were detected by UHPLC-Q-TOF-MS~E technique, and the obtained mass spectrometry data was combined with UNIFI software for prediction. The prototype components and metabolites in rat feces were identified with reference materials and related literature. A total of 43 metabolites were identified(including 8 prototype components and 35 metabolites). The metabolic pathways mainly include monocaffeoylquinic acid(hydrogenation reduction, ring-opening cracking, sulfation, hydroxylation, glucuronidation), quercetin(O-C2 bond ring-opening cleavage, C2-C3 double bond reduction, rutin carbonylation) and so on. The metabolites and metabolic process of L. bulbifera extract in rat feces were clarified, which provided a basis for the study of the active substances and its mechanism of action.

Diagnosis and treatment of intrahepatic cholangiocarcinoma / 国际外科学杂志

Intrahepatic cholangiocarcinoma (ICC) is a liver malignant tumor, and its incidence is second only to hepatocellular carcinoma, accounting for 5%-30% of patients with primary liver cancer. Usually it is characterized by insidious attack, high degree of malignancy, postoperative recurrence, and being asymptomatic at the early stage, most patients have developed into the advanced stage when the diagnosis is confirmed. Thus, early laboratory examinations and imaging judgments are particularly important, providing opportunities for early radical resection of patients. The purpose of this paper is to review and update the medical literature to provide new ideas for diagnosis and treatment of ICC.

Ater-soluble Protein Profiles of Astragalus membranaceus var. mongholicus and A. membranaceus / 中国实验方剂学杂志

Objective::Astragalus membranaceus var. mongholicus and A. membranaceus are medicinal Astragalus, which are closely related and similar in composition, but with unclear medicinal value. Water-soluble protein profiles for A. membranaceus var. mongholicus and A. membranaceus were established to explore the differences between the two kinds of Astragalus Radix. Method::The water-soluble protein components were obtained through water ultrasonic extraction and acetone precipitation. After digested with trypsin, the obtained peptides were analyzed by nano ESI-LC-MS/MS method. Proteome Discovery 1.4 software was used to identify the proteins by comparing with the legume protein database, and the different expression water-soluble proteins were analyzed by the label-free quantitative software SIEVE. Finally, relevant information for common expression proteins, including classification, molecular function, involved biological process and signaling pathway, were analyzed by bioinformatics. Result::There were 920 and 717 specific proteins identified for A. membranaceus var. mongholicus and A. membranaceus, respectively. Totally 472 proteins were found to be co-expressed, in which 21 were differentially expressed, such as PR-10 protein, NDK-1 protein, glutelin A2, and phospholipase D. There were 14 highly expressed proteins in A. membranaceus var. mongholicus and 7 highly expressed proteins in A. membranaceus. Conclusion::There are significant differences in water-soluble protein profiles for two kinds of Astragalus Radix. Specific proteins, differentially expressed proteins and common expressed proteins can provide references for the identification of A. membranaceus var. mongholicus and A. membranaceus. It also can be used to define pharmacological mechanisms and search for drug action targets.

Value of lymph node ratio in prognosis of patients with intrahepatic cholangiocarcinoma / 中华肝胆外科杂志

Objective To determine the value of lymph node ratio (LNR) in prognosis of patients with intrahepatic cholangiocarcinoma (ICC).Methods The pathological and survival data of ICC patients were extracted from the Surveillance Epidemiology and End Result Database (SEER).The value of LNR in prognosis of patients with intrahepatic cholangiocarcinoma was evaluated by the Kaplan-Meier and Cox proportional hazard models.Results A total of 357 pathologically confirmed ICC patients were included in this study (158 males,199 females).The results showed that patients with lymph node metastasis had significantly shorter survival time than patients without metastasis [(41.6 ± 1.8) vs.(23.2 ± 2.2) months,P＜ 0.05].Multivariate analysis on survival showed that the status of lymph node metastasis was independent prognostic factor of survival,after adjusting for age,gender and tumor stage.Only a LNR ≥ 20％ was an independent risk factor of prognosis (HR=2.3,95％CI:1.4～3.8,P＜0.05).Conclusion A positive lymph node ratio was superior to lymph node metastasis in predicting survival in patients with intrahepatic cholangiocarcinoma.

Comparison of in Vitro Dissolution of Active Ingredients in Gastrodiae Rhizoma Powder with Different Particle Size / 中国实验方剂学杂志

Objective: To compare in vitro dissolution behaviors of active ingredients (gastrodin,parishin A,p-hydroxybenzyl alcohol,parishin B and parishin C) in Gastrodiae Rhizoma powder with different particle size.Method: In vitro dissolution of Gastrodiae Rhizoma powder in different dissolution media (water,artificial gastric juice and artificial intestinal juice) were detected by stirring paddle method.Dissolution of these five components in Gastrodiae Rhizoma powder with different particle size was determined by HPLC,mobile phase was acetonitrile-0.1% phosphoric acid solution for gradient elution,column temperature was set at 40℃ and detection wavelength was 220 nm.Result: In water and artificial intestinal juice,the dissolution rates of five active components in three kinds of Gastrodiae Rhizoma ultrafine powders were higher than that of the fine powder and the finest powder;in artificial gastric juice,the dissolution rates of gastrodin and p-hydroxybenzyl alcohol in Gastrodiae Rhizoma ultrafine powders were higher than that of the other powders,and the dissolution rate of parishin A in Gastrodiae Rhizoma ultrafine powders was lower than that of the other powders.Conclusion: An appropriate degree of superfine grinding can promote the dissolution of active ingredients in Gastrodiae Rhizoma,but not as fine as possible.The dissolution medium has an obvious influence on the dissolution behaviors of active components,which provides a reference for screening optical particle size of Gastrodiae Rhizoma powder in clinical application.

Protective Effect of Gastrodiae Rhizoma Powder with Different Particle Sizes on Cerebral Ischemia Reperfusion Injury in Rats / 中国实验方剂学杂志

Objective: To study the protective effect of Gastrodiae Rhizoma ultrafine powder with different particle sizes on focal cerebral ischemia-reperfusion injury in rats, in order to provide theoretical basis for the application of ultrafine powder technology in related products. Method: The SD rats were randomly divided into six groups, namely sham-operated group, ischemia-reperfusion group, Gastrodiae Rhizoma ultramicro-powder Ⅰ group(1.2 g·kg-1), Gastrodiae Rhizoma ultramicro-powder Ⅱ group(1.2 g·kg-1), Gastrodiae Rhizoma ultramicro-powder Ⅲ group(1.2 g·kg-1), and nimodipine positive drug group(12.6 g·kg-1). The cerebral ischemia-reperfusion model was established after 7 days of drug administration. The focal ischemia reperfusion injury model was prepared by middle cerebral artery occlusion (MCAO) for 1 h followed by reperfusion for 24 h. The infarct size of brain tissue was measured by triphenyltetrazolium Chloride(TTC) staining. The activities of superoxide dismutase (SOD), malondialdehyde (MDA) and nitricoxide synthase (NOS) in brain homogenate and serum were measured by colorimetry. The contents of glutamate(Glu), aspartic acid(Asp) and glycine(Gly) in hippocampus were determined by liquid chromatography-tandem mass spectrometry (LC/MSMS). Result:The model group showed obvious signs of neurobehavioral deficit (PPPPPConclusion:Gastrodiae Rhizoma ultramicro-powder with different particle sizes has a protective effect on cerebral ischemia-reperfusion injury, and the Gastrodiae Rhizoma ultramicro-powder Ⅲ group has a more obvious protective effect than the the Gastrodiae Rhizoma ultramicro-powder Ⅰ group. The results showed that the protective effect of Gastrodiae Rhizoma ultramicro-powder on middle cerebral artery ischemia-reperfusion in rats was related to its particle size.

Serum Pharmacochemical Analysis of Periploca forrestii Rhizomes Based on UPLC-Q-TOF-MS / 中国实验方剂学杂志

Objective:To study the serum pharmacochemistry of Periploca forrestii rhizomes,and to investigate the pharmacological material basis of extract of P. forrestii rhizomes in rats. Method:Rapid identification of constituents absorbed into blood was carried out by UPLC-Q-TOF-MS,according to retention time,accurate relative molecular mass and standard substance comparison,these constituents were identified and speculated by Data Analysis,Metabolite Detect and other softwares,then preliminary determination of constituents absorbed into blood of rats after oral administration of extract of P. forrestii rhizomes was investigated. Result:Totally 17 constituents absorbed into blood were detected in serum,ten of them were prototype constituents and the other were metabolites.Seven of the prototypes were identified as 5-O-caffeoylquinic acid,4-O-caffeoylquinic acid,3-O-caffeoylquinic acid,3,4-di-O-caffeoylquinic acid,3,5-di-O-caffeoylquinic acid,4,5-di-O-caffeoylquinic acid and periplocin. Conclusion:These constituents absorbed into blood may be substances that act directly in vivo of P. forrestii rhizomes,and it is helpful to clarify pharmacological material basis and mechanism of this herb.

Kinetic Analysis of Enzymatic Reaction of Active Components from Xinshao Formula in Liver Microsomes of Sham-Operated and Cerebral Ischemia-reperfusion Injury Rats / 中国实验方剂学杂志

Objective: To investigate and compare enzymatic kinetics of scutellarin,apigenin-7-O-glucronide and paeoniflorin from Xinshao fomula in liver microsomes of sham-operated rats and middle cerebral artery occlusion(MCAO) rats with focal cerebral ischemia-reperfusion injury. Method: Xinshao fomula were incubated respectively with liver microsomes of sham-operated rats and MCAO rats,UPLC-MS and substrate elimination method was employed,Michaelis constant(Km),maximum velocity of enzymatic reaction(Vmax) and intrinsic clearance(CLint) of these three components from Xinshao fomula in liver microsomes of sham-operated rats and MCAO rats were calculated,these parameters between different groups were evaluated by statistical analysis. Result: The Km values of scutellarin,apigenin-7-O-glucronide and paeoniflorin in liver microsomes of MCAO rats were (0.798±0.031),(0.213±0.017),(0.499±0.029) μmol·L-1,which were quite different to these in liver microsomes of sham-operated rats.Compared with the sham-operated group,Vmax and CLint values of scutellarin and paeoniflorin in liver microsomes of MCAO rats were significantly reduced(PPVmax of apigenin-7-O-glucronide in liver microsomes of MCAO rat was also significantly reduced(PConclusion: Metabolic rates of these three active components from Xinshao fomula in liver microsomes of MCAO rats with focal cerebral ischemia-reperfusion injury decrease with low elimination rate.

Determination of plasma protein binding rates of nine compounds of Inula cappa extraction based on method of equilibrium dialysis / 中国中药杂志

To determine the plasma protein binding rate of the nine compounds in Inula cappa extraction by the method of equilibrium dialysis. The proteins in plasma samples were precipitated by methanol, and the ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was developed for determination of the concentrations of the nine active compounds, namely chlorogenic acid, scopolin, neochlorogenic acid, cryptochlorogenic acid, 1,3-O-dicaffeoylquinic acid, galuteolin, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, with the internal standard of puerarin. We found that all components have a good linearity(r≥0.999), and accuracy, precision, extraction recovery and stability conformed to the requirements of determination, without endogenous compounds disturbing within the range of optimum concentration. This suggested that the method was stable and reliable, and could be used for the determination of the plasma protein binding rates of the nine active compounds in rat and human plasma of I. cappa. The plasma protein binding rates of the nine active compounds in rat and human plasma respectively were(41.07±0.046)%-(94.95±0.008)%, and(37.66±0.043)%-(97.46±0.013)%. According to the results, there were differences in the plasma protein binding rates of the nine compounds in I. cappa extraction between rat and human.