Objective: To investigate the mechanism of berberine regulating glycogen metabolism and its effect on glycogen structure. Methods: The db/db mice were used as diabetic mice to investigate the effect of berberine on blood glucose levels in db/db mice. The liver glycogen was extracted for SEC and TEM analysis to investigate the effect of berberine on liver glycogen and the mechanism of its effect. Results: Berberine significantly decreased the fasting blood glucose level and the insulin level in serum of db/db mice, and berberine repaired the damaged glycogen structure. Meanwhile, berberine decreased the expression of GP, GDBE, cAMP in liver tissue and the glucagon level in serum of db/db mice. Conclusion: Berberine regulated the cAMP/GP signaling pathway, improved hepatic glycogen structure in db/db mice, and repaired damaged glycogen structure, which may be one of the mechanisms for regulating hepatic glucose metabolism and improving symptoms of diabetes.

Objective: To prepare the taxifolin and determine its apparent oil-water partition coefficient in different media, and to study the mechanism of absorption and transport of taxifolin in Caco-2 cell model. Methods: Taxifolin was prepared by enzymolysis. HPLC was used to determine the saturated solubility of taxifolin in 37 ℃, different pH buffer solution and water, apparent oil-water distribution coefficient of taxifolin obtained by calculation formula of oil-water distribution coefficient; CCK-8 experiment was used to investigate the safe concentration range of taxifolin in Caco-2 cells, and then the single-layer model of Caco-2 cells was used to study the mechanism of bilateral transmembrane absorption and transport. CCK-8 experiment was used to investigate the safe concentration range of taxifolin in HDMEC cells. The inflammatory model of HDMEC cells induced by lipopolysaccharide was established, and the activity of lactic dehydrogenase was detected by the intervention of floxacin. The activity of lactic dehydrogenase was detected by lactic dehydrogenase kit. Results: The lgP values of taxifolin in the following solvents were 0.29 (0.1 mol/L hydrochloric acid), 0.48 (pH 2.0), 0.46 (pH 5.8), 0.34 (pH 6.8), 0.26 (pH 7.4), and 0.38 (water), respectively; There was no significant toxic effect on Caco-2 cells in the range of 50-500 μg/mL; There was no significant difference in Papp value of bilateral transport between different concentrations of taxifolin in Caco-2 monolayer cell model, and it was less than 1 × 10-6 cm/s and ER was less than 2. There was no significant toxic effect on HDMEC cells in the range of 50-300 μg/mL; After treatment with taxifolin, compared with LPS stimulation group, the activity of LDH in each treatment group was decreased significantly (P < 0.05), and the activity of LDH was decreased significantly in the range of 50-100 μg/mL, and tended to be stable in the range of 100-250 μg/mL. Conclusion: Taxifolin is a kind of drug which is difficult to absorb in the intestine. The mechanism of transmembrane transport is passive transport. It can inhibit the inflammation of hdmec cells induced by LPS and has anti-inflammatory activity.

Influenza is a common infective respiratory system disease in pediatric clinical. Based on the guidelines for diagnosis and treatment of influenza in children at home and abroad, guidelines for clinical trials of influenza drugs, and related registrations or published clinical trials combined with clinical experiences, the key technical points and characteristics of clinical trial design and evaluation of Chinese medicine for this disease was mainly elaborated, in order to enrich the methodological content of traditional Chinese medicine clinical evaluation of pediatric diseases.

Objective: It is difficult to accurately grasp the essential characteristics of medicinal properties of traditional Chinese medicine due to the abstraction and vagueness. This paper proposes a Quantitative Model of Traditional Chinese Medicine's Properties based on BP Neural Network (QM-BP Model) to train and realize quantitative representations of Chinese herbal medicine (CHM). Methods: Data for analysis were obtained and organized by conceptual analysis. Sample pairs of the associations were obtained based on the relationships of CHM and their efficacy. Then a QM-BP model with three-tier structure in form of CHM-drug vector-efficacy was constructed, initialized and trained according to prior organized CHM data. Finally, rules of correlation of CHM and their efficacy was obtained by training dataset with drug vectors representing quantitative attributes of CHM. Results: Based on the training of QM-BP model, 474 TCM and 528 effects included in the textbook of TCM were trained and combined based on the training of QM-BP model. It was found that the BP drug vectors representing drug properties after training reflected the attribute characteristics of CHM better than the initial quantitative values. In addition, as BP drug vector and word vector have similar properties, the BP drug vectors for CHM with similar efficacy was relatively close in Euclidean distance while the CHM with different efficacies were relatively far in Euclidean distance. Conclusion: In this paper, a BP neural network was adopted to construct a medicine vector training model. Based on the correlation between the medicinal properties and efficacy of TCM, the quantified values of the medicinal properties were modified to represent medicinal properties more accurately. In future work, the QM-BP model can be applied to the analysis of herb pairs and prescriptions to analyze the rules of combination related to medicinal properties and the compatibility within prescriptions.

Objective: To explore the prescription medication rule of the first batch of Chinese medicine masters in treating acute stage of stroke by using the data mining method. Methods: The experience books and published journal articles of Chinese medicine masters and the shared medical records of national famous TCM studios in the cloud platform V1.5 of ancient and modern medical records were searched; The first batch of national physician master data for the treatment of acute phase of stroke was selected; A standardized basis database, standardization of medicines were established after using statistical analysis of system integration, association rules analysis, methods of complex networks; The formula frequency, medicinal, and core prescription drugs were analyzed. Results: A total of 142 medical cases were eventually included. The frequency results of traditional Chinese medicine showed that 14 kinds of high-frequency traditional Chinese medicine were obtained, including Pheretima, Chuanxiong Rhizoma, Persicae Semen, etc., and most of the drugs were warm, smooth, bitter, sweet, and return to liver and lung meristem. The results showed that the frequency of chordal pulse and tongue red was the most. A total of 24 association rules and 18 TCM association rules were obtained by association rule analysis. Conclusion: In the treatment of acute apoplexy, the Chinese medicine master usually adopted the methods of clearing heat and relieving wind, promoting blood circulation and removing blood stasis, eliminating phlegm and resuscitative medicinal, moving qi, and dredging collaterals, and stopping convulsions.

Objective: To clone AbCYP80F1 gene and its promoter, analyze the Cis-acting elements related to control signals and transcription factors in AbCYP80F1 gene promoter and study the effect of AbCYP80F1 overexpression on scopolamine accumulation in Atropa belladonna. Methods: CYP80F1cDNA from Hyoscyamus niger was used as query sequence to do blastn search in A. belladonna transcriptome database to retrieve homologous unigenes. Full-length of AbCYP80F1 cDNA and gene promoter were acquired by RACE and hiTAIL-PCR, respectively. Agrobacterium rhizogenes-dipping method was adopted to induce AbCYP80F1- overexpressed A. belladonna hairy roots, and scopolamine content in hairy roots was detected by HPLC. Results: A full-length of 1 675 bp of AbCYP80F1 gene was obtained and the deduced protein had a closed homology with CYP80F1 from Anisodus luridus. A length of 2 000 bp promoter contained Cis-acting elements related to light, anaerobic induction, auxin, MeJA, gibberellins and low temperature, as well as recognition sites of transcription factor of MYB, AP2/ERF, WRKY and bHLH. Overexpression of AbCYP80F1 gene increased the scopolamine content in hairy roots of A. belladonna by an average of 1.8 times. Conclusion: AbCYP80F1 is a secondary root-specific gene in scopolamine biosynthetic pathway and its overexpression can enhance scopolamine accumulation. Expression of AbCYP80F1 may be regulated by WRKY and bHLH transcription factors.

Objective: To obtain the functional genes in Arctium lappa and analyze the key enzyme genes involved in biosynthesis pathway of lignin. Methods: The transcriptome dataset of roots of A. lappa was obtained by using the BGISEQ-500 sequencing platform. Unigenes were de novo assembled and annotated according to the existing nucleic acids and protein databases. The key enzyme genes involved in lignin biosynthesis pathway were analyzed and the three-dimensional model of phenylalanine ammonialyase (AlPAL) was generated by the SWISS-MODEL and PyMol. Results: A total of 54 215 Unigenes were obtained by de novo assembly, and 42 003 Unigenes were annotated in at least one public database. A total of 1 668 Unigenes were identified to be plant transcription factors (TFs), which belong to 54 TF families, and 423 Unigenes were found to be involved in the biosynthesis pathway of lignin. Structure model indicated that AlPAL was a homotypic tetramer, and each monomer was consisted of three domains, including 4-methyl-imidazole-5-ketone (MIO) domain, core domain and shield domain. The MIO domain contained three conserved amino acids (ASG), which formed the catalytic activity center of the enzyme. Conclusion: This study was the first de novo transcriptome assembly of A. lappa, which will lay the foundation for the identification of functional genes, secondary metabolic pathway and the study of regulation mechanism of biosynthesis pathway of lignin in A. lappa in the future.

Objective: To develop the EST-SSR molecular identification system of Lilium lancifolium, Lilium davidii var. willmottiae, Lilium regale, Lilium casa blanca and Lilium brownie var. viridulum, and analyze the development efficiency and identification ability of EST-SSR molecular marker technology for Lilium genus. Methods: The MISA.pl program was used to identify the SSR locus of the EST gene sequence published by NCBI. The EST-SSR primers were generated by Primer3 program module, and the primers were screened by PCR amplification and agarose gel electrophoresis. The primary screening primers were verified by polyacrylamide gel electrophoresis, and the characteristic bands of different germplasms were labeled and analyzed to construct an identification system. Results: A total of 199 pairs of SSR primers were designed. After screening 26 pairs of primers, two pairs of highly efficient primers were obtained. The molecular identification system constructed by primer JZ391 can effectively identify the mixed commercial materials, which had certain practical value. Conclusion: Based on the extreme genetic characteristics of the research materials, this identification marker development is very efficient. The results confirm that the genetic basis of the species is an important factor affecting the development efficiency of its EST-SSR molecular marker. At the same time, this case can be used as a reference for the development of EST-SSR markers for other Chinese medicinal herbs similar to Lilium genus.

Objective: To analyze and identify the significant different components between Cordyceps hawkesii and Cordyceps sinensis by using method of ultra performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS). Methods: Mass spectrometry combined with formula finder of PeakView software and database (Human Metabolome Database, Pub Chem, Metlin) and secondary fragmentation analysis, significant different components were identified and analyzed. Results: Through OPLS-DA analysis, it was found that 12 significant different components were identified. Eleven of them were amino acids and their metabolites, and one was phosphatidylcholine. Conclusion: Surprisingly, characteristic components such as cordycepin and adenosine were not identified by significant difference analysis. In this study, it was proved that C. hawkesii can be used as a supplementary resource instead of C. sinensis, which provided scientific support for the further development and utilization of C. hawkesii.

Objective: To establish a rational method for Laportea bulbifera quality control. Methods: The fingerprint technique and multi-component quantitation were used to study the quality control of L. bulbifera by UHPLC. The 12 batches of L. bulbifera UHPLC fingerprint were evaluated by the evaluation system on similitude degree of chromatogram fingerprint of traditional Chinese medicine. Results: The quality control methods of Miao medicine L. bulbifera for simultaneous determination of 10 components (including neochlorogenic acid, cryptochlorogenic acid, chlorogenic acid, rutin, isoquercitrin, luteoloside, kaempferol-3-O-rutinoside, quercitrin, epigallocatechin and epicatechin) were established. The linear, precision, repeatability and stability are good. The standard recoveries were 95.89%-98.62%, with RSD less than 3%. The common mode of fingerprint was established after determination fingerprints of 12 batches of samples of L. bulbifera by UHPLC. There were 20 common peaks in these samples. The similarity of the 12 batches fingerprints were in the range from 0.805 to 0.931. Conclusion: The fingerprinting and multi-index content determination methods for quantitative control of Miao medicine L. bulbifera have high sensitivity, good accuracy, stability and reliability, which can provide a theoretical and experimental foundation for quantitative control of Miao medicine L. bulbifera.