Study on tanshinones regulating root-associated microbiomes of Salvia miltiorrhiza / 中国中药杂志

The plant root-associated microbiomes include root microbiome and rhizosphere microbiome, which are closely related to plant life activities. Nearly 30% of photosynthesis products of plants are used to synthesize root compounds, there is evidence that root compounds regulate and significantly affect the root microbiome Tanshinones are the main hydrophobic components in Salvia miltiorrhiza. In order to study whether these compounds can regulate the root-associated microbiomes of S. miltiorrhiza, our study first identified a white root S. miltiorrhiza(BG) which contains little tanshinones. Retain of the fifth intron of tanshinones synthesis key enzyme gene SmCPS1 leading to the early termination of the SmCPS1 gene, and a stable white root phenotype. Further, wild type(WT) and BG were planted in greenhouse with nutrient soil(Pindstrup, Denmark) and Shandong soil(collected from the S. miltiorrhiza base in Weifang, Shandong), then high-throughput sequencing was used to analyze the root-associated microbiomes. The results showed that the tanshinones significantly affected the root-associated microbiomes of S. miltiorrhiza, and the impact on root microbiomes was more significant. There are significant differences between WT and BG root microbiomes in species richness, dominant strains and co-occurrence network. Tanshinones have a certain repelling effect on Bacilli which belongs to Gram-positive, while specifically attract some Gram-negative bacteria such as Betaproteobacteria and some specific genus of Alphaproteobacteria. This study determined the important role of tanshinones in regulating the structure of root-associated microbiomes from multiple angles, and shed a light for further improving the quality and yield of S. miltiorrhiza through microenvironment regulation.

Analysis of Rhizosphere Soil Fungal Community Composition and Diversity of Salvia miltiorrhiza Based on ITS Sequences / 中国实验方剂学杂志

Objective: To study rhizosphere soil fungal community composition and diversity of Salvia miltiorrhiza,in order to explain the changes of the rhizosphere microenvironment of S.miltiorrhiza,and provide theoretical basis for revealing the reasons for the formation of continuous obstacles.Method: Based on the high-throughput sequencing technology,three kinds of soil samples were collected,namely soil of non-plant (CK),soil for cultivating for one year (Y1) and soil for cultivating for two years (Y2),respectively.According to the sequencing of the internal transcribed space of nuclear ribosomal DNA (ITS) region of fungal rRNA gene,14 153,19 024 and 7 622 valid sequences were obtained and annotated as 1 027,1 095 and 712 varieties of OTUs.The diversity of fungal communities showed a trend of decreasing first and then increasing.Agaricomycetes,Leotiomycetes and Tremellomycetes were the dominant bacterial communities at the class level,their orders in three treatments were Y1 > CK > Y2,Y2 > Y1 > CK,CK > Y2 > Y1.At the genus level,Guehomyces was dominant species in CK,and both Y1 and Y2 had a decrease of 96.13% and 90.21%,respectively.Wickerhamomyces had a significantly increase in Y2(6.64%),250.59% and 564.00% compared with CK and Y1,respectively.Result: The results of cluster analysis showed that CK and Y1 had a higher similarity at the genus level in soil fungal community composition compared with Y2.Conclusion: Cryptosporiopsis and other pathogenic fungal communities were dominant genuses according to the heatmap analysis in Y2,suggesting a correlation with the incidence of S.miltiorrhiza.The findings provide theoretical basis for the formation of continuous cropping obstacle.

Influence of naturaldecay of fibrousroots on growth and content of active compositions of Salvia miltiorrhiza f. alba / 中国中药杂志

The change of yield and contents. of active compositions were studied while the fibrous roots were decayed naturally. HPLC method was used to detect the contents of active composition. The results show that fibrousroots could decrease the production of plant by 38.60% (20 g) and 30.99% (40 g), respectively. Treatment 1 could increase the contents of dihydrotanshinone and cryptotanshinone of Salvia miltiorrhiza f. alba by 26.08% and 22.64%, respectively. Compared with the comparison, treatment 2 decreased the contents of ihydrotanshinone, cryptotanshinone, tanshinone I and tanshinone II(A) of S. miltiorrhiza f. alba by 60.87%, 79.24%, 84.61% and 88.99%, respectively. Meanwhile, the total contents of the liposoluble constituents reduced by 86.27%. The different concentration of fibrousroots could increase the content of salvianolic acid B by 4.98% (20 g) and 23.64% (40 g), respectively. Meanwhile, the content of rosemary acid was increased by 4.98% (20 g) and 23.64% (40 g), respectively. The content of water-soluble constituents positively correlated to the mount of added fibrousroots, and the change was significantly. The result indicted that the decay of fibrousroots has a significant impact on the growth and the content of the active composition of S. miltiorrhiza f. alba under the condition of continuous cropping. Fibrousroots could decrease the content of biomass and liposoluble constituents significantly, which maybe one of the main factors to S. miltiorrhiza f. alba continuous cropping obstacle formation.

Influence of continuous cropping years on yield and active compounds in Salvia miltiorrhiza f. alba / 中国中药杂志

Biological characteristic of Salvia miltiorrhiza f. alba in field was studied. HPLC method was used to determine the lipophilic constituents (dihydrotanshinone, cryptotanshinone, tanshinone, tanshinone II (A) and miltione) and hydrophilic constituents (salvianolic acid, rosemarinic acid). The results showed that the fresh weight of S. miltiorrhiza f. alba which cropped for 2 years was decreased by 80.47%, while dry weight decreased by 79.42%. The normal diameter of the root was 0.3-0.5 cm, however, the diameter was 0.2-0.4 cm after 2 years, it was said that the decrease of the root diameter was the main reason for the decrease of the yield. The average contents of dihydrotanshinone, cryptotanshinone, tanshinone, tanshinone II (A), miltione, salvianolic acid and rosemarinic acid were decreased by 35.26%, 32.26%, 19.35%, 3.39%, 64.40%, 66.93% in plant which continuously cropped for 2 years, respectively. The yield and active constituents were mostly effected in the plant of S. miltiorrhiza f. alba, which continuously cropped for 2 years.

S100A4 siRNA inhibits human pancreatic cancer cell invasion in vitro / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Pancreatic cancer is one of the most deadly cancers, which is characterized by its high metastatic potential. S100A4 is a major prometastatic protein involved in tumor invasion and metastasis which precise role in pancreatic cancer has not been fully investigated. We knocked down the S100A4 gene in the Bxpc-3 pancreatic cancer cell line via RNA interference to study the changes in cell behavior.</p><p><b>METHODS</b>Real-time polymerase chain reaction and western blotting were used to detect mRNA and protein expression levels of S100A4, matrix metalloproteinase (MMP)-2, E-cadherin and thrombospondin (TSP)-1. Transwell chambers were used to detect the migration and invasion abilities; a cell adhesion assay was used to detect adhesion ability; colony forming efficiency was used to detect cell proliferation; flow cytometry was used to detect apoptosis.</p><p><b>RESULTS</b>S100A4 mRNA expression was reduced to 17% after transfection with S100A4-siRNA, and protein expression had a similar trend. mRNA and protein expression of MMP-2 was reduced and that of E-cadherin and TSP-1 was elevated, indicating that S100A4 affects their expression. S100A4-silenced cells exhibited a marked decrease in migration and invasiveness and increased adhesion, whereas overall proliferation and apoptosis were not overtly altered.</p><p><b>CONCLUSION</b>S100A4 and its downstream factors play important roles in pancreatic cancer invasion, and silencing A100A4 can significantly contain the invasiveness of pancreatic cancer.</p>

Primary research of anti-influenza virus activity of "Benovoair Concentrate" / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Anti-influenza virus activity of "Benovoair Concentrate".</p><p><b>METHODS</b>The different dilution of samples were mixed with the same quantity of 100 TCID50 virus at 37 degrees C for 30 minutes. Add suitable quantity mixture in wells containing cells. Every 3 wells were the same mode. Viruses control, cells control and samples control of different dilution were performed and set in the CO2 incubator at 37 degrees C. CPE was observed every day. When CPE appears in viruses control as "++++", stopped testing and performed the hemagglutination titration.</p><p><b>RESULTS</b>"Benovoair Concentrate" with dilution of 1:60, 1:120, 1:240 and 1:480 have 100% anti-influenza A and anti-influenza B activities. "Benovoair Concentrate" with dilution of 1:960 and 1:1920 have 25%-50% anti-influenza A and anti-influenza B activities.</p><p><b>CONCLUSION</b>The test was the proof of anti-influenza virus activities which provided for the development of "Benovoair Concentrate".</p>