Exploration on Syndrome Types and Metabolic Markers of Rat Model with Heart Failure Caused by Transverse Aortic Constriction Based on Theory of Correspondence of Prescription and Syndrome / 中国实验方剂学杂志

ObjectiveTo observe the difference in the efficacy of three kinds of traditional Chinese medicine （TCM） injections on rat model of heart failure induced by transverse aortic constriction （TAC）， explore the TCM syndrome of the model based on the theory of correspondence of prescription and syndrome， and reveal the biological basis of prescription-syndrome from the perspective of metabolism. MethodRats were treated with TAC for modeling and were divided into Shenmai injection group （6.0 mL·kg-1）， model group， Danhong injection group （6.0 mL·kg-1）， Shenfu injection group （6.0 mL·kg-1） and trimetazidine group （10 mg·kg-1）， and sham operation group was set up as control. After drug intervention for 15 days， echocardiography， serum N-terminal pro-brain natriuretic peptide （NT-proBNP） and myocardial histopathological staining were performed for each group， so as to compare the efficacy to select the effective injection. Colorimetry was used to detect the serum glucolipid metabolism after the intervention of the effective injection， and ultra high performance liquid chromatography-mass spectrometry was used to observe the metabolites and related metabolic pathways in myocardial tissue. ResultCompared with the sham operation group， the left ventricular ejection fraction （LVEF） and left ventricular fractional shortening （FS） in the model group decreased （P<0.01）， while the left ventricular end-diastolic diameter （LVIDd）， left ventricular internal diameter at end-systole （LVIDs） and NT-proBNP level increased （P<0.01）. Compared with model group， LVEF and FS increased （P<0.01）， LVIDd， LVIDs and NT-proBNP level decreased （P<0.05， P<0.01） in Danhong injection group， NT-proBNP level in Shenfu injection group decreased （P<0.05）， LVIDd and NT-proBNP level increased （P<0.05， P<0.01） in Shenmai injection group， in trimetazidine group， LVEF and FS increased （P<0.01）， while LVIDs and NT-proBNP level decreased （P<0.05， P<0.01）. Serum glucose， low-density lipoprotein cholesterol and high-density lipoprotein cholesterol levels in Danhong injection group and trimetazidine group were adjusted by callbacks （P<0.01， P<0.05）. There were the callback of 9 myocardial metabolites in Danhong injection group， including glycine， serine and threonine metabolism， glyoxylate and dicarboxylate metabolism， glycerol phospholipid metabolism. There were the callback of 10 myocardial metabolites in trimetazidine group， including glycerol phospholipid metabolism. ConclusionThe efficacy of Danhong injection on heart failure model induced by TAC is significant and superior to Shenfu injection and Shenmai injection， suggesting that the model is closely related to heart-blood stasis. The biological mechanism of Danhong injection interfering with the model involves regulating the metabolic disorder of lipid， glucose， amino acid and butyric acid.

Improvement of Quality Control Standard of Bufonis Venenum in Shexiang Baoxin Pills / 中国实验方剂学杂志

ObjectiveTo develop a quantitative analysis of multi-components by single marker （QAMS） for determination of bufalin， cinobufagin and resibufogenin in Shexiang Baoxin pills， and to provide a method for improving the national standard of the pills. MethodHigh performance liquid chromatography （HPLC） was developed for simultaneous determination of bufalin， cinobufagin and resibufogenin in Shexiang Baoxin pills and the methodology validation was carried out. The chromatographic separation was performed on a Nucleosil 100-5 C18 column （4.6 mm×250 mm， 5 μm） with the mobile phase of acetonitrile -0.1% potassium dihydrogen phosphate aqueous solution （pH adjusted to 3.2 with phosphoric acid） （48∶52）， and the flow rate was 0.6 mL·min-1， the detection wavelength was set at 296 nm and the column temperature was 35 ℃. Taking cinobufagin as the internal standard， the relative correction factors （RCFs） of bufalin and resibufogenin were calculated， and the key influencing factors of RCFs were investigated. Relative retention time was used for the chromatographic peak location of the analyte， combining with the on-line ultraviolet spectroscopy and accurate relative molecular weight obtained by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-QTOF/MS）. The external standard method was used to verify the contents of three components obtained by QAMS. ResultQAMS was established for the determination of bufalin， cinobufagin and resibufogenin in the samples， and RCFs of cinobufagin to bufalin and resibufogenin were 0.922 and 1.01， respectively. The total content of the three marker compounds in 11 batches of Shexiang Baoxin pills was 33.7-36.0 µg per pill. There was no significant difference between the quantitative results of QAMS and external standard method. ConclusionThe established method can be used for the quality control of bufalin， cinobufagin and resibufogenin in Shexiang Baoxin pills. It is suggested that bufalin should be considered as one of three marker compounds， and the sum of bufalin， cinobufagin and resibufogenin should be used for the content limit of this preparation.

Pathological Process Observation and Non-targeted Metabolomics Analysis of Heart Failure Rat Model Established by Transverse Aortic Constriction / 中国实验方剂学杂志

ObjectiveTo study the pathological process and changes of metabolites in myocardial tissue of heart failure induced by transverse aortic constriction （TAC） in rats. MethodRats were treated with TAC operation and divided into TAC-30 d group and TAC-60 d group， and sham operation group at the same period was set up as control. Echocardiography and pathological staining of myocardial tissue were performed on rats in each group. Enzyme-linked immunosorbent assay was used to determine the expression of amino-terminal pro-brain natriuretic peptide （NT-proBNP） and adenosine triphosphate （ATP） in serum. Liquid chromatography-mass spectrometry was used to observe the changes of metabolites and related pathways in myocardial tissue， the mobile phase consisted of 25 mmol·L-1 ammonium acetate and 25 mmol·L-1 ammonia hydroxide in water （A） and acetonitrile （B） for gradient elution （0-0.5 min， 95%B； 0.5-7 min， 95%-65%B； 7-8 min， 65%-40%B； 8-9 min， 40%B； 9-9.1 min， 40%-95%B； 9.1-12 min， 95%B）， electrospray ionization was used under positive and negative ion detection modes， acquisition range was m/z 70-1 050. ResultCompared with the sham-30 d group， the left ventricular internal diameter at end-systole （LVIDs） in TAC-30 d group was significantly decreased （P<0.01）， and left ventricular ejection fraction （LVEF）， fraction shortening （FS）， left ventricular end-diastolic posterior wall thickness （LVPWd）， left vebtricular end-systolic posterior wall thickness （LVPWs） were significantly increased （P<0.01）， there were cardiomyocyte arrangement disorder， edema， collagen fibre hyperplasia， the content of NT-probNP was significantly increased， while the content of ATP was significantly decreased （P<0.01）， and 15 metabolites with abnormal expression were involved in pyrimidine metabolic pathway， pantothenic acid and coenzyme A biosynthesis pathway. Compared with the sham-60 d group， LVEF and FS in the TAC-60 d group were significantly decreased （P<0.01）， and left ventricular internal diameter at end-diastole （LVIDd）， LVIDs and LVPWd were increased （P<0.05， P<0.01）， the edema of myocardial cells increased obviously， myocardium fibers degenerated， coagulation necrosis appeared， and a large amount of collagen fibers were deposited， the expression of NT-proBNP increased and the expression of ATP decreased （P<0.01）， there were 21 metabolites with abnormal expression， involving pyrimidine metabolic pathway， and starch and sucrose metabolic pathway. ConclusionAt 30 d after TAC， there are myocardial hypertrophy， lipid metabolism disorder， pyrimidine metabolism disorder and energy imbalance. At 60 d after TAC， there are heart failure， aggravation of lipid metabolism disorder， excessive activation of glucose metabolism， and continuous disorder of pyrimidine metabolism.

Effects of Huatan Jiangzhuo decoction on diet-induced hyperlipidemia and gene expressions in rats / 中国天然药物

Huatan Jiangzhuo decoction (HJD) is a combination of six traditional Chinese medicines that were used for lipid metabolism-related disorders, but its efficacy and underlying mechanisms have not been explored by modern research strategies. This study aimed to investigate the therapeutic role of HJD in determining the transcriptome level. Hyperlipidemia model was established by feeding Sprague-Dawley rats with high-fat diet. Differentially expressed genes (DEGs) were detected by high-through transcriptome sequencing, followed by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The total cholesterol (TC) and triglyceride (TG) levels in hyperlipidemia model rats were significantly increased, whereas high-density lipoprotein (HDL) concentration decreased when compared to normal rats, and HJD significantly downregulated TC concentrations and liver coefficient in the hyperlipidemia rats. Histology staining showed that HDJ greatly recovered the lipid accumulation in rat hepatic stellate cells and aortic arch vascular wall thickness of hyperlipidemia rats. One thousand nine hundred and thirty-six DEGs were identified in the HJD-treated hyperlipidemia rats, which were associated with various biological processes and signaling pathways such as peroxisome proliferator-activated receptors, AMP-activated Protein Kinase , and insulin signaling pathways. Quantitative reverse transcription-polymerase chain reaction further confirmed the downregulated expression of cholesterol 7-α-hydroxylase(CYP7A1), liver orphan receptor(LXRα),peroxisome proliferator-activated receptor gamma(PPARγ),andSterol Response Element-Binding Protein 1c(SREBP1c) genes in hyperlipidemia rats treated with HJD. Our data first elucidated the gene expression profile of high-fat diet-induced hyperlipidemia in rats after HJD treatment, and lipid metabolism-related genes (CYP7A1, LXRα, PPARγ, and SREBP1c) may be potentially biomarkers for HJD-alleviated hyperlipidemia.

Separation and structural elucidation of C25 epimers of furostanol saponin from aerial parts of Paris polyphylla var. chinensis / 中国中药杂志

Paris polyphylla var. chinensis(PPC) is used as one of the origin plants of Paridis Rhizoma described in the Chinese Pharmacopoeia(2020 edition). Its resources shortage makes the planting scale gradually expand, and plenty of aerial parts are abandoned because of not being effectively used. On the basis of previous research, this study separated steroidal saponins to further clarify the chemical composition of the aerial parts of PPC. As a result, three pairs of 25R or 25S epimers of furostanol saponins were obtained by various column chromatography techniques. Their structures were identified as neosolanigroside Y6(1), solanigroside Y6(2), neoprotogracillin(3), protogracillin(4), neoprotodioscin(5) and protodioscin(6) by spectral data combining with chemical transformation. Compound 1 is a new compound, and compounds 2, 3 and 5 are isolated from Paris plants for the first time. Compounds 4 and 6 are isolated from this plant for the first time. Previously, only several spirostanol glycosides with 25S configuration were isolated from Paris plants. Guided by mass spectrometry, the present study isolated the furostanol saponins with 25S configuration from this genus for the first time, which further enriches the chemical information of Paris genus and provides a reference for the isolation of similar compounds.

Rapid discovery of tyrosinase inhibitors from Sophora flavescens:significance for comprehensive utilization of its non-medicinal resources / 中国中药杂志

Sophorae Flavescentis Radix,derived from the root of Sophora flavescens in the Leguminosae family,has been widely used in the medicine,agriculture,animal husbandry,and daily chemical industry. A pharmacophore model-based method for rapid discovery of tyrosinase inhibitors( TIs) from S. flavescens was established by molecular docking under Lipinski rules,and verified by enzyme assays. Briefly,the chemical constituent database of S. flavescens( CDSF) was established based on the previous papers. Theoptimal pharmacophore model( OPM) was constructed by DS 2019 on the basis of known active TIs. Eighty-three hits predominated by flavonoids having higher fitting scores with OPM than the positive control were screened out,and subjected to molecular docking based on the three-dimensional structure of tyrosinase crystal protein. The potential TIs such as kurarinone and nor-kurarinone were rapidly discovered from the compounds with higher docking scores than the positive control under the Lipinski rules. The results were verified by the in vitro enzyme assays. The inhibition activities of tyrosinase from non-medicinal parts of S. flavescens were also tested to explore the relationship between the inhibition activity and chemical compositions. This study is expected to provide data support for the comprehensive application and development of S. flavescens and also a new method for the rapid discovery of active substances or functional constituents in the complex systems.

Application of QAMS for quality evaluation and control of Chinese patent medicines:taking Bufonis Venenum-contained preparations as examples / 中国中药杂志

As a new strategy capable of uncovering the characteristics of traditional Chinese medicines, the quantitative analysis of multi-components by single-marker(QAMS) has been widely employed for the quality evaluation of Chinese medicinal materials, slices, and extracts. However, its application in the assessment of Chinese patent medicines is yet to be explored. By referring to the determination of three bufogenins in Bufonis Venenum by QAMS described in Chinese Pharmacopoeia(2020 Edition), this paper selected seven representative preparations containing Bufonis Venenum and explored whether the relative correction factors(RCFs) of cinobufagin(CB) to bufalin(BF) and resibufogenin(RB) could be directly used for the quality control of Bufonis Venenum-contained preparations. Based on the qualitative analyses under the same chromatographic conditions as used for toad venom, combing specificity test, five preparations such as Yatong Yili Pills, Houzheng Pills, Xiongdan Jiuxin Pills, Liushen Pills and Niuhuang Xiaoyan Pills, were expected to use validated RCFs for the direct determination of three components. Taking Houzheng Pills as an example, the methodological validation of bufalin, cinobufagin and resibufogenin was carried out, and the recoveries of bufalin, cinobufagin and resibufogenin were 90.64%-106.1%. The obvious difference was not observed between the contents of bufalin and resibufogenin in 24 batches of preparation samples by QAMS and external reference method. In the tested samples, the content of bufalin, cinobufagin and resibufogenin were 1.27-2.61, 2.44-5.66 and 0.988-3.16 mg·g~(-1) in 10 batches of Liushen Pills samples. The contents of bufalin, cinobufagin and resibufogenin were 0.760-1.32, 1.35-2.39 and 0.600-1.55 mg·g~(-1) in 10 batches of Houzheng Pills samples from three manufacturers. The obtained data contribute to improving the quality standard of Bufonis Venenum-contained preparations, and they also provide some ideas for the application of QAMS in the quality evaluation and control of Chinese patent medicines.

Improving Quality Standard of Processed Slices of Paridis Rhizoma of Chinese Pharmacopoeia / 中国实验方剂学杂志

Objective:To study the appearance description,TLC examination and content determination was carried out, in order to improve the quality standard of processed slices of Paridis Rhizoma in the 2020 edition of Chinese Pharmacopoeia. Method:Based on the literature review and observation on the samples,the appearance description was described. TLC examination was used for the qualitative analysis. HPLC was used for the determination of polyphyllin Ⅰ,Ⅱ,Ⅵ and Ⅶ in the commercial and processed samples. UPLC was employed for the determination of 10 steroidal saponins,namely pseudoprotodioscin,polyphyllin Ⅶ,17-hydroxygracillin,polyphyllin H,polyphyllin Ⅵ,polyphyllin Ⅱ,dioscin,gracilin,polyphyllin Ⅰ and polyphyllin Ⅴ. Result:For the appearance description,color and luster,texture,odor and taste as well as the diameter of 1.0-4.5 cm were recorded. polyphyllin Ⅵ was not detected in the thin layer chromatograms of most of the tested samples derived from high-quality species but obviously detected in those of Trillium Rhizoma. Five of 13 commercial samples met the requirements that the total amounts of polyphyllin Ⅶ,Ⅵ,Ⅱ,and Ⅰ should be no less than 0.6%according to the current Chinese Pharmacopoeia. Because softening and drying had the obvious influence on the contents of steroidal saponins in the samples,soaking and sun-drying were preferred. Conclusion:Appearance description should be supplemented. Polyphyllin Ⅵ is not considered as one of quality markers for the TLC identification and HPLC determination of Paridis Rhizoma. Polyphyllin H was considered as a new marker for the quality control. It is recommended that the total amounts of polyphyllin Ⅶ,H,Ⅱ,and Ⅰ should be no less than 1.0%.

Aspirin interrupts bile duct carcinoma in rats induced by thioacetamide / 第二军医大学学报

Objective To establish a thioacetamide (TAA)-induced bile duct carcinoma rat model and to observe the effect of aspirin on bile duct carcinoma by interrupting the model. Methods Sprague-Dawley (SD) male rats were given drinking water containing TAA (300 mg/L) for 5 months, and tumor formations of the livers were observed by H-E staining at the 8th, 12th, 16th, and 20th weeks, respectively. The expression of carbonic anhydrase 2 (CA-2) in liver tissues was also detected by immunohistochemistry. SD male rats were also selected and treated with aspirin after exposing to TAA for 12 weeks. And then the tumor formations of the livers were observed by H-E staining at the 3rd and 6th months, and the expression of CA-2 was detected by immunohistochemistry. Bile duct carcinoma cells QBC939 were obtained and treated with aspirin (0 mmol/L and 5 mmol/L), and the expression of CA-2 was detected by Western blotting after culturing for 48 h. Results Obvious fibrosis was found in the livers of some rats at 12th weeks after exposing to TAA; large number of fibrous tissue hyperplasia and microscopic or suspected tumors were found in the livers at the 16th weeks; visible tumors in the livers were found in all the rats at the 20th weeks. The incidence of hepatic tumors or suspected tumors was 28.6% (2/7) in rats treated with aspirin for 6 months, while it was 100% (7/7) in rats without aspirin treatment. The expression of CA-2 in liver tissues was gradually increased with the development of bile duct carcinoma in rats, while the expression of CA-2 in liver tissues was decreased after treating with aspirin. The expression of CA-2 in QBC939 cells treated with 5 mmol/L aspirin for 48 h was significantly decreased versus the untreated control group (P0.01). Conclusion TAA can successfully induce intrahepatic bile duct carcinoma. Aspirin can interrupt the development of bile duct carcinoma and may be used to prevent intrahepatic bile duct carcinoma.

Study on quality control method of toad venom based on characteristic chromatogram and QAMS / 中国中药杂志

Toad venom (Chansu) is prepared from the dried secretion of parotid gland and skin gland from Bufo bufo gargarizans or B. melanostictus. Up to now, much attention shall be paid to the poor quality of commercial toad venom because of the adulteration. So, it is urgent to establish a scientific and perfect quality control method to improve the quality of toad venom and guarantee its safety and effectiveness in clinical application. The different batches of toad venom samples were assayed by high performance liquid chromatography (HPLC) and the quantitative analysis of multi-components by single marker (QAMS) was used to detect the contents of five bufagenins. As a result, the reference characteristic chromatogram was established, displaying serotonin, gamabufotalin, arenobufagin, hellebrigenin, telocinobufagin, bufotalin, cinobufotalin, bufalin, cinobufagin and resibufogenin as characteristic peaks. Taking cinobufagin as an internal reference substance, QAMS was verified for the determination of five bufagenins (gamabufotalin, bufotalin, bufalin, cinobufagin, resibufogenin) in toad venom samples. The durability and applicability of the relative correction factor (RCF) were also studied systematically. RCFs of cinobufagin to gamabufotalin, bufotalin, bufalin and resibufogenin were determined as 1.05, 0.895, 1.09 and 0.913, respectively. The characteristic chromatogram and QAMS established in this study could effectively control the quality of toad venom and provide scientific evidence for the improvement of the quality standard of the toad venom to be described in Chinese Pharmacopoeia (2020 edition).

Effects of Early Administration of Dexmedetomidine on Benzodiazepine Requirements of ICU Patients with Alcohol Withdrawal Syndrome / 中国药师

Objective:To evaluate the effects of early administration of dexmedetomidine on benzodiazepine ( BZD) requirements of intensive care unit ( ICU) patients with alcohol withdrawal syndrome ( AWS) .Methods:Totally 60 adult patients with severe AWS were chosen and randomly divided into groups A and group B with 30 patients in each .The patients in group A received benzodiazepine simultaneously with early administration of dexmedetomidine , and those in group B received only benzodiazepine .After the first admin-istration of dexmedetomidine , the 24-h cumulative BZD requirement , stay length in ICU and hospital , condition of mechanical ventila-tion and incidence of hypotension and bradycardia were recorded .Results:The 24-h cumulative BZD requirement in group A was obvi-ously lower than that in group B [(8.6 ±1.2) mg vs (25.6 ±4.9) mg, P<0.05].The number of patients with mechanical ventila-tion in the two groups was similar, while the duration of intubation in group A was less than that in group B [(24.9 3.1) h vs (48.8 6.9) h, P<0.05].Additionally, group A had higher incidence of bradycardia when compared with group B (57% vs 13%, P<0.05).Conclusion:As the adjuvant treatment for alcohol withdrawal syndrome , early administration of dexmedetomidine can notably decrease the benzodiazepine requirement of ICU patients .

Development of the Chinese age norms of CES-D in urban area / 中国心理卫生杂志

Objective: To test applicability of the Center for Epidemiological Studies Depression Scale (CES-D) in different age groups in urban China, and to develop age norms. Methods: In this cross-sectional study, the CES-D was administrated to 16047 community participants with average age of (37.7±21.3) years (age ranged 11～100) in 21 provinces, who were as the norming sample. Criterion validity was tested in 349 psychiatric patients with average age of (32.0±12.1) years (age ranged 16～81) in 4 cities. A subsample (199 workers, 100 col-lege students, and 30 teachers in Beijing, Dongguan, and Baotou) was drawn from the national sample to provide 8 week interval test-retest reliability. Results: The Cronbach α was 0.90 for the scale, and 0.68～0.86 for its fac-tors. The 8 week interval test-retest correlation was 0.49 for the scale (P <0.01) and 0.39～0.51 for factors (P<0.01) . The result of confirmatory factor analysis supported the original 4-factor structure (RMSEA=0.057, CFI =0.976, GFI=0.948) . Patients scored higher than community sample [(21.72±13.39 ) vs.(13.24±10.33),P <0.01], and depression patients scored the highest [(27.82±14.42), P<0.01] . Age difference was signifi-cant. Age groups over 60-year-old scored higher than all the other age groups under 60-year-old (P<0.01). Con-clusion: The Chinese version of CES-D shows good refiability and validity across all ages in urban population-