ABSTRACT
italic>Glycyrrhiza eurycarpa P.C.Li is a medicinal plant resource and is often mixed with traditional licorice herbs. We sequenced the chloroplast genome of Glycyrrhiza eurycarpa P.C.Li using Illumina high-throughput sequencing technology, and physical mapping and genomic characterization was carried out. Comparative genomic analysis was performed with Glycyrrhiza uralensis Fisch, Glycyrrhiza inflata Bat and Glycyrrhiza glabra L. The Glycyrrhiza eurycarpa P.C.Li chloroplast genome was 127 864 bp long with 34.25% GC content, consisting of a large single copy and a small single copy. The genome was missing the inverted repeat (IR) region. A total of 110 genes were annotated, including 76 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. The 301 SSRs, rich in A-T repeats, were detected by MISA. The Glycyrrhiza eurycarpa P.C.Li chloroplast genome showed weak codon preference, and the codons were biased to use A and T bases. Three specific gene fragments of Glycyrrhiza eurycarpa P.C.Li were characterized by homology comparison. Based on Pi analysis, six new high mutation regions (psbZ-psbC, trnC-GCA-rpoB, trnR-UCU-trnG-UCC, ycf2, trnN-GUU-ycf1, ndhA) of medicinal licorice species were determined. The results of phylogenetic analysis indicate that Glycyrrhiza eurycarpa P.C.Li from Xinjiang is an interspecific hybrid taxon closely related to the three medicinal licorice species, and Glycyrrhiza inflata Bat, which is distributed in the same domain, is its male parent. Based on this study, the taxonomic identification, herb-specific DNA fingerprint development, genetic diversity, and molecular plant breeding of medicinal plants of the genus Glycyrrhiza can be established.
ABSTRACT
Thin layer chromatography, high performance liquid chromatography and multivariate statistical analysis were integrated in current study to provide a basis for the quality evaluation and the standard improvement of Paridis Rhizoma(Chinese name: Chong-lou). The results demonstrated that the primary saponins in the two authorized sources of Paridis Rhizoma were polyphyllinsⅠ, Ⅱ and Ⅶ, while the rhizome of Trillium tschonoskii an adulterant of Paridis Rhizoma was rich of polyphyllin Ⅵ. Therefore, the apparent content of polyphyllin Ⅵ plays a determinant role towards the source authentication of raw materials and decoction slices of Paridis Rhizoma, whose adulterants frequently occur in the market. Moreover, the contents of polyphyllin Ⅵ in the two authorized sources could meet the requirements of Chinese Pharmacopoeia. Therefore, we suggested that polyphyllin Ⅵ should not be omitted from the quality standard of Paridis Rhizoma in the Chinese Pharmacopoeia, and on the other side, polyphyllinsⅠ, Ⅱ and Ⅶ should be the eligible quality indicators. The study aims to sound information and evidences for the quality evaluation of Paridis Rhizoma, and also to provide a theoretical basis for the standard revision of Paridis Rhizoma in the future Chinese Pharmacopoeia.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Rhizome , Saponins , TrilliumABSTRACT
The content of tyrosol,salidroside,echinacoside,rutin,acteoside,ligustroflavone,specnuezhenide,and quercetin were determined by HPLC,and the color of Ligustri Lucidi Fructus was determined by comparison with color card.Hundred-seed weight was analyzed by using gravimetric method.The correlation analysis and One-way ANOVA were used to analyze the relationship between the characters,the chemical composition,the harvest time and the geographical location of Ligustri Lucidi Fructus,for giving a comprehensive evaluation of the quality of Ligustri Lucidi Fructus The results showed that 92% of Ligustri Lucidi Fructus were all up to quality standard of Chinese Pharmacopoeia,and the contents of 7 components in Ligustri Lucidi Fructus(except quercetin) were higher than those in samples with black colors.The content of salidroside in Ligustri Lucidi Fructus harvested in June was the highest and the other7 components of Ligustri Lucidi Fructus were relatively high in 8-10 months.According to the quality parameters of Ligustri Lucidi Fructus,the Ligustri Lucidi Fructus from six habitats can not be distinguished effectively.The results showed that there was a certain relationship between the color,harvest season and component content of Ligustri Lucidi Fructus,and the habitats were not related to the quality parameters of Ligustri Lucidi Fructus.The study aimsat providing data support for the resource status of native Ligustri Lucidi Fructus,and a theoretical basis for the revision of standards of Ligustri Lucidi Fructusin the future.
Subject(s)
Drugs, Chinese Herbal , Reference Standards , Fruit , Chemistry , Ligustrum , ChemistryABSTRACT
Objective: To establish the method for the specific chromatograms analysis of steamed Panax notoginseng Powder and determine the content of eight components, combined with clustering analysis and partial least square discriminant analysis (PLS-DA), with aim to provide reference for the quality control of steamed Panax notoginseng Powder. Methods: HPLC with Agilent ZORBAX SB-C18 column (250 mm × 4.6 mm, 5 μm) was used, the mobile phase was acetonitrile (B)-water (A) in a gradient elution mode, the detection wavelength was set at 203 nm, the flow rate was 1.0 mL/min, the column temperature was 40 ℃, and sample size 10 ìL. HPLC characteristic spectrum of steamed Panax notoginseng Powder was established, and quantitative determination methods of eight index components, ginsenoside 20 (S)-Rh1, 20 (R)-Rh1, Rk3, Rh4, 20 (S)-Rg3, 20 (R)-Rg3, Rk1 and Rg5, were investigated, its content in 63 batches of samples were determined. Results: The specific chromatograms of steamed Panax notoginseng Powder effective parts were established and 19 common peaks were designated. Among them, eight rare saponins including ginsenoside 20 (S)-Rh1, 20 (R)-Rh1, Rk3, Rh4, 20 (S)-Rg3, 20 (R)-Rg3, Rk1 and Rg5 all showed good linear relationship within the ranges of 0.999 9, 0.999 5, 0.999 4, 0.999 3, 0.999 1, 0.999 3, 0.999 1, and 0.999 3, respectively. The average recovery was 95%—105%, with the RSD value less than 2%. Moreover, the 63 batches of samples were divided into two groups: brown red and light yellow, and the quality of brown red group was obviously better than that of pale yellow group. Conclusion: The quality of the steamed Panax notoginseng Powder is related to the color and lustre. This method is accurate, sensitive and reproducible, which can provide reference for the quality evaluation of steamed Panax notoginseng Powder.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of embryonic stem cells on the proliferation and apoptosis in human acute myeloid leukemia cell line KG-1a and to explore its potential mechanism.</p><p><b>METHODS</b>The direct co-culture system between human embryonic stem cells H9 and human acute myeloid leukemia cell line KG-1a was established, and CCK8 assay was used to detect the proliferation of KG-1a cells. The changes of cell cycle and apoptosis were detected by flow cytometry (FCM). The mRNA expressions of BCL-2, BAX, Caspase-3 were assessed by RT-PCR. Meanwhile, the protein-expressions of BCL-2, BAX, Caspase-3 were detected by Western blot.</p><p><b>RESULTS</b>The proliferation level of KG-1a cells was significantly inhibited by H9, and the apoptotic rate increased, and the cell cycle was blocked at G/M phase. The mRNA-expression and the protein-expression of BAX and Caspase-3 increased, the mRNA and protein-expression of BCL-2 decreased.</p><p><b>CONCLUSION</b>Embryonic stem cells can inhibit the proliferation of KG-1a and induce the apoptosis that maybe relate with the down-regulation of BCL-2 expression and up-regulation of BAX and caspase-3 expression.</p>
Subject(s)
Humans , Apoptosis , Caspase 3 , Cell Line, Tumor , Cell Proliferation , Human Embryonic Stem Cells , Leukemia, Myeloid, Acute , Proto-Oncogene Proteins c-bcl-2 , bcl-2-Associated X ProteinABSTRACT
This study was designed to investigate the inhibitory effect of supernatant from co-culture of human embryonic stem cells and tumor MDA-MB-231 cells on the breast cancer. The direct co-culture system of human embryonic stem cells H9 and breast cancer MDA-MB-231 cells was established, and the supernatant was tested in the inhibition of MDA-MB-231 cells. The inhibitory effects were examined in tumor cell morphology using microscope, cell proliferation with MTT assay, and cell apoptosis using the Hoechst staining and flow cytometry. Transwell assay was used to detect the migration and invasion of tumor cells. The results suggest that the supernatant significantly inhibited the proliferation, invasion and migration, and promoted cell apoptosis of MDA-MB-231 cells. However, the supernatant of H9 cells alone had little effect on MDA-MB-231 cells. Therefore, we conclude that the supernatant of co-culture cells had an inhibitory effect on tumor cells in vitro.