Research progress in correlation between commodity specification level and quality of Polygala tenuifolia / 中草药

The quality of Chinese herbal medicine is an important prerequisite for ensuring clinical efficacy and safety. The current commodity specification of medicinal herbs is the summary of the traditional Chinese medicine practitioners' experience of "assessing the quality by distinguishing the features of traditional Chinese medicinal materials". However, with the continuous introduction and acclimatization and large-scale cultivation of wild medicinal herbs, the traits of some medicinal herbs have changed greatly compared with the previous one, resulting in that the market circulation specifications of these medicinal herbs is inconsistent with the current commodity specification. In this paper, by consulting the modern monographs, literatures and so on, we analyzed the market circulation specifications being implementing of Polygalae Radix, combined with the modern quality evaluation methods to explore the effects of strains and base, origin and climate, growth duration and harvesting period and medicinal parts on the quality of P. tenuifolia. This paper reveals the correlation between the traits and quality of Polygalae Radix, furthermore, provides the theoretical basis for setting up the commodity specifications of Polygalae Radix, and determines the future research direction of P. tenuifolia quality assessment.

Analysis of influencing factors of secondary metabolites contents in cultivated Polygala tenuifolia / 中国中药杂志

This work was launched to explore the effect of habitat and growth year on the secondary metabolites contents of cultivated Polygala tenuifolia. The samples of cultivated P. tenuifolia were analyzed by ultra-high performance liquid chromatography(UPLC)-quadrupole time-of-flight mass spectrometry(Q-TOF MS), and the obtained data were analyzed using multiple statistical analysis and cluster analysis. The results showed that compared with growth year, habitat is a main influencing factor which affected the secondary metabolites contents of P. tenuifolia. The contents of sucrose esters and oligosacchride multi-esters are greatly dependent on the habitat (the sample-AG with high levels of components of tenuifoliside B and tenuifoliside C, and the sample-FY with high levels of 3,6'-disinapoyl sucrose, tenuifoliose S, tenuifoliose L, and tenuifoliose V). There is no obvious effect of habitat and growth year on xanthone. The contents of triterpene saponins are greatly dependent on the growth year, and the content of parts of triterpene saponins increased as time goes on.The result indicated that the effect of habitat and growth year on different types of secondary metabolites is not completely equivalent. This study will contribute to the breeding of P. tenuifolia and amendment of current commodity criteria.

Analysis of digital gene expression profiles of the cultivated Polygala tenuifolia in different phenological phases / 药学学报

The content changes of chemical components in different phenological phase of the cultivated Polygala tenuifolia is one of the important factors for determination of the best harvest time in the production practice. In this study, the digital gene expression (DGE) profiles of the cultivated P. tenuifolia were analyzed in different phenological phase (flowering fruit bearing stage, wilting stage, dormancy stage). The differentially expressed genes were found in the biosynthesis of chemical composition in P. tenuifolia, and the representational ones were validated by RT-qPCR. Then, the key enzymes (CYP450s and UGTs) involved in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia were predicted through the correlation analysis of gene expression. The number of down-regulated genes was more than that of up-regulated in P. tenuifolia from flowering fruit bearing stage to dormancy stage. Six differentially expressed genes (HMGS, PMK, FPPS, SQS, SE, β-AS) and five (PAL, C4H, 4CL, CAD, peroxidase) were annotated to the triterpenoid saponins and phenylpropanoid biosynthesis pathway in P. tenuifolia, respectively. Compared to wilting and dormancy stages, the saponins, xanthones, and lignins were largely synthesized at the flowering fruit bearing stage of P. tenuifolia. Furthermore, UGT83A1, CYP716B1, CYP98A3, CYP86B1, and CYP94A1 may be the part of key enzymes in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia. This study provides evidence to support the correctness of traditional harvest time of P. tenuifolia at the level of transcription, and lays the scientific foundation for gene cloning and functional verification of CYP450s and UGTs in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia in the future.

Rationality of commodity criteria and traditional breeding of Polygala tenuifolia based on agronomic traits and determination of chemical components / 中国中药杂志

The agronomic traits (plant height, root diameter, root length, first lateral root height, lateral root amount, root weight) of 18 Polygala tenuifolia samples with different agronomic traits were analyzed, respectively. HPLC was used to analyze three main characteristic components including tenuifolin, polygalaxanthone Ⅲ, and 3,6'-disinapoyl sucrose. At last, the correlation between six agronomic traits and three main characteristic components were analyzed by scatter plot. We found no significant correlation between root diameter and three main characteristic components. There were no obvious correlations between tenuifolin and the remaining five agronomic traits. Short root length and first lateral root height as well as high lateral root amount resulted in high levels of polygalaxanthone Ⅲ in P. tenuifolia samples. High levels of 3,6'-disinapoyl sucrose were observed in P. tenuifolia samples with longer root. So, the current commodity criteria and traditional breeding of P. tenuifolia did not conform to pharmacopoeia standards, which excellent medicinal materials should have high contents of the main characteristic components. It was urgent to revise the current commodity criteria and breeding methods.

Comparison between Astragalus membranaceus var. mongholicus and Hedysarum polybotrys based on ITS sequences and metabolomics / 药学学报

Astragalus membranaceus var. mongholicus and Hedysarum polybotrys belong to different genera, but have similar drug efficacy in traditional Chinese medicine theory, and H. polybotrys was used as the legal A. membranaceus var. mongholicus previously. In this study, similarities and differences between them were analyzed via their ITS/ITS2 fragments information. The ITS (internal transcribed spacer) regions were amplified using polymerase chain reaction and then sequenced in two-way. The alignment lengths of ITS regions were 616 bp, in which 508 loci were consistent, and 103 loci were different, accounting for 82.47% and 16.72% of the total ITS nucleotides in length, respectively. As genotype determines phenotype, 1HNMR-based metabolomic approach was further used to reveal the chemical similarities and differences between them. Thirty-four metabolites were identified in the 1H NMR spectra, and twenty-seven metabolites were the common components. Amino acids, carbohydrates and other primary metabolites were similar, while a large difference existed in the flavonoids and astragalosides. This study suggests that A. membranaceus var. mongholicus and H. polybotrys show similarities and differences from molecular and chemical perspectives, which has laid a foundation for elucidating the effective material basis of drug with similar efficacy and resources utilization.

UPLC/Q-TOF MS and NMR plant metabolomics approach in studying the effect of growth year on the quality of Polygala tenuifolia / 药学学报

Growth year is one of the important factors for the quality of Polygala tenufolia. In this study, primary metabolites and secondary metabolites were compared in 1, 2 and 3 years old P. tenufolia cultivated in Shaanxi Heyang. The samples were subjected to ultra-high performance liquid chromatography (UPLC)-quadrupole time-of-flight mass spectrometry (Q-TOF MS) and nuclear magnetic resonance (NMR) analysis, and the obtained data were analyzed using principal component analysis (PCA) and other statistical analysis methods. In addition, content and correlation of different metabolites were also calculated. The results showed no significance between main component contents in 2 year-old and 3 year-old P. Tenufolia, but 1 year-old was statistically different. The contents of primary metabolites, such as fructose, sucrose, and choline increased as time goes on, while glycine and raffinose decreased. The contents of secondary metabolites, such as onjisaponin Fg, polygalasaponin XXVIII, polygalasaponin XXXII increased, while polygalaxanthone III and parts of oligosaccharide multi-ester including tenuifoliose A, tenuifoliose C, tenuifoliose C2 and tenuifoliose H decreased with the extension of the growth years. Growth years has important impact on the quality of P. tenuifolia and the existing growing years of commodity P. tenuifolia have its scientific evidence. This study supplied a new method for the quality evaluation of Chinese medicinal materials.

Between Hengshanhuangqi and Chuanhuangqi based on metabolomics and ITS2 sequences / 药学学报

To compare the differences between Hengshanhuangqi (HH) and Chuanhuangqi (CH) at molecular level, 1H NMR based plant metabolomics approach was used to reveal the chemical difference between HH and CH. Then, the contents of astragaloside IV and calycosin-7-O-beta-D-glucoside, the marker compounds specified in China Pharmacopoeia, were determined. In addition, the ITS2 fragments of HH and CH were sequenced. Twenty-three metabolites were identified in the 1H NMR spectrum, and the principal component analysis showed CH and HH could be separated clearly. HH contained more aspartic acid, GABA, citric acid, astragaloside IV and calycosin-7-O-beta-D-glucoside, while CH contained more threonine, alanine, acetic acid, choline, arginine, fructose and sucrose. And the astragaloside IV is almost undetectable in CH. In addition, the ITS2 fragment sequences of HH and CH were different at eight bases. Thus, the HH and CH showed significant differences chemically and genetically.

Establishment and optimization of ISSR reaction system for Anoectochilus formosanus / 中草药

Objective: To establish a stable, reproducible, and suitable reaction system of Anoectochilus formosanus for ISSR analysis of genetic differences according to the ISSR-PCR characters in plants of Anoectochilus Bl. Methods: The initial ISSR reaction system in plants of Anoectochilus Bl. was established and then the primers were screened. After that, PCR amplifications were carried out until the different concentrations of the factors in the ISSR reaction system were designed, and based on the principle of high stability and reproducibility, the stable and reliable ISSR reaction system was eventually established after a series of adjustment and optimization of the reaction system. Results: The optimal ISSR reaction system in plants of Anoectochilus Bl. was reported for the first time, and the volume of 25 μL contained approximately 20 ng template DNA, 1 U Taq DNA polymerase, 0.2 mmol/L deoxyribonucleotide triphosphate (dNTP), 2 mmol/L MgCl2, 200 umol/L dNTP Mixture, and 0.2 μmol/L primer within 1x reaction buffer [10 mmol/L Tris-HCl (pH 8.3), 50 mmol/L KCl]. The reaction mixtures were denatured at 94°C for 7 min and subjected to 45 cycles of 30 s at 94 °C, 45 s at 52 °C, 2 min at 72 °C, and a final extension step of 10 min at 72 °C and eventually stored at 4 °C. Conclusion: The ISSR-PCR systems established for studying on the plants of Anoectochilus Bl. show the characters of stable reaction system, better repeatability, clear marker site, and reliable abundant polymorphisms. It is suitable for the study on genetic diversity in plants of Anoectochilus Bl. in difference wild populations and genetic variation of its sterile seedlings after long time tissue culture.

Study on the mechanism of scar formation: epidermis template defect theory / 中华烧伤杂志

Dermal defection and the degree of its loss determine the natural process of wound healing, which is the key reason leading to excess scar hyperplasia. The function of tri-dimensional structure in dermis acts as a template to regulate the properties of reparative cells. The template structure induces the reparative cells to grow into the structure which changes the skin mechanic status on wound area. Also, the component of extracellular matrix can affect behaviours of fibroblasts negatively or positively, for the reason that the structure of dermal tissue has a permissive effect on the dermal components in regulating behaviours of reparative cells. Therefore, the behaviors of cells depend on the structure of the template. The suitable tri-dimensional structure of dermis facilitates normal cell cycling. The more the structure of dermis closed to its physiological status, the better the biological behaviors of cells act. Moreover, the integrity as well as the continuity of dermal tissue is the prerequisite for serving as a template. The damage to the integrity and the continuity of dermal tissue may be one of the key reasons to lead abnormal tissue repair and scar formation. Thus, we hypothesize that the loss of dermal template may be one of the mechanism of abnormal scar formation and propose the theory of extracellular matrix framework deficiency or destruction.

Mild hypothermia reduces and delays cell apoptosis in the hippocampal CA_1 region following ischemia-reperfusion injury / 中华物理医学与康复杂志

Objective To investigate the protective effects of mild hypothermia on cells in the CA_1 region of the hippocampus in gerbils following global ischemia-reperfusion injury(IRI),and to explore their mechanism. Methods IRI models were established in 75 gerbils.Any changes in TUNEL positive cells and the expression of Bax and Cytochrome C were then observed in normothermic(N) ,hypothermic(H)and sham(S) groups through immu- nohistochemistry methods.Results In the H group(as compared with the N group)apoptotic cells in the CA_1 sub- field of the hippocampus significantly decreased.The expression of Bax and Cyt C at 3 h,6 h and 1 d were de- creased,and the expression fastigium was delayed.Conclusion Mild hypothermia can moderate and delay cell ap- optosis,and its mechanism might be related with reducing and delaying the expression of Bax and Cyt C released by mitochondria.