OBJECTIVETo study the patterns of dynamic accumulation of total flavonoids and total saponins in the roots of Ophiopogon japonicus collected from different harvest times, and compare the contents of total flavonoids and total saponins in roots of O. japonicus which were processed with different methods.

METHODThe total flavonoids and total saponins contents in O. japonicus were determined by ultraviolet spectrophotometry.

RESULT AND CONCLUSIONFrom December to January, the total contents of flavonoids and saponins in the roots of O. japonicus gradually decreased, and gradually increased from February to March, and kept stable in April. The contents of total flavonoids and total saponins in the O. japonicus were influenced by different processing methods.

Desiccation ; Flavonoids ; analysis ; metabolism ; Ophiopogon ; chemistry ; metabolism ; Plant Roots ; chemistry ; metabolism ; Saponins ; analysis ; metabolism ; Seasons ; Temperature ; Time Factors

OBJECTIVETo develop an HPLC-ELSD method for the determination of 25 (R, S) ruscogenin 1-O-[beta-D-glucopyranosyl (1 --> 2)] [beta-D-xylopyranosyl (1 --> 3)] beta-D-fucopyranoside in the tuberous roots of Liriope muscari from different habitats and different harvest time.

METHODA Shimadzu C18 column (4.6 mm x 150 mm, 5 microm) with a solvent system consisting of acetonirile-water (46: 54) was used, and detected by ELSD. The temperature of drift tube was 94 degrees C and the nebulizer nitrogen flow rate was 2.5 L x min(-1).

RESULTThe calibration curve of 25 (R, S) ruscogenin 1-O-[beta-D-glucopyranosyl (1 --> 2)] [beta-D-xylopyranosyl (1 --> 3)] beta-D-fucopyranoside showed good linearity in the range of 1.02-12.228 microg and the average recovery was 100.80%, with RSD of 1.8%. 10 batches of L. muscari from different habitats were analyzed, and the contents were 0.25% - 0.41%. The contents of 15 batches from different harvest time were 0.13%-0.38%.

CONCLUSIONThe method is simple, rapid and sensitive, and can be used for determination of 25 (R, S) ruscogenin 1-O-[beta-D-glucopyranosyl (1 --> 2)] [beta-D-xylopyranosyl (1 --> 3)] beta-D-fucopyranoside in L. muscari. It provides the valuable basis for quality assessment of L. muscari.

Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Liliaceae ; chemistry ; Liriope Plant ; chemistry ; Magnetic Resonance Spectroscopy ; methods ; Molecular Sequence Data ; Molecular Structure ; Plant Preparations ; analysis ; chemistry ; pharmacology ; Plant Roots ; chemistry ; physiology ; Plant Structures ; chemistry ; Saponins ; chemistry ; Spirostans ; analysis ; chemistry ; pharmacology ; Triterpenes ; isolation & purification

Background: Anti-programmed death-1/programmed death-ligand 1 (PD-1/PD-L1) immunotherapy has been proved to be effective on gastric cancer in ongoing clinical trials. However, the value of PD-L1 in predicting responses of patients with gastric cancer to anti-PD-1/PD-L1 immunotherapy is controversial. Some studies suggested that intra- and inter-tumoral heterogeneity of PD-L1 expression might explain the controversy. This study aimed to analyze the expression of PD-L1, PD-L2, and PD-1 as well as CD8(+) T-cell density in primary tumors and lymph nodes from patients with stage T1-4N+M0 gastric adenocarcinoma to explore the heterogeneity of PD-1 signaling pathway molecules. Methods: In primary tumors and metastatic as well as non-metastatic lymph nodes from patients with stage T1-4N+M0 gastric adenocarcinoma, we detected PD-L1 and PD-L2 expression with immunohistochemistry. CD8(+) T-cell density in primary tumors and PD-1 expression on CD8(+) T cells were detected with immunofluorescence. Uni-variate analysis was used to determine the prognostic values of them. Cox proportional hazard regression model was used to identify independent risk factors that affect patients' overall survival and disease-free survival. Results: Among 119 eligible patients who had undergone surgical resection, the positive rate of PD-L1 was higher in metastatic lymph nodes than in primary tumors (45.4％ vs. 38.7％,P= 0.005); the positive rate of PD-1 on CD8(+) T cells was significantly higher in primary tumors and metastatic lymph nodes than in tumor-free lymph nodes (both P < 0.001). The intensity of PD-1 expression on CD8(+) T cells in primary tumors and in metastatic lymph nodes were stronger than that in tumor-free lymph nodes from the same patient. Beside, the positive rate of PD-L2 did not show any differences between primary tumors and metastatic lymph nodes. In multivariate analysis, PD-L1 expression, PD-L2 expression, a low density of CD8(+) T cells in primary tumors, and PD-1 expression on CD8(+) T cells in primary tumors were associated with poor prognosis.Conclusion: The expression of PD-L1 is heterogeneous in primary tumors and in metastatic lymph nodes from patients with stage T1-4N+M0 gastric adenocarcinoma, which might explain the inconsistent results in assessing the prognostic value of PD-L1 expression in previous studies.

OBJECTIVE To investigate the effects of different drying methods on the index components in wine-processed Cornus officinalis so as to optimize drying method.METHODS After processed with wine， C. officinalis decoction pieces were dried with different drying methods （blast drying， far infrared drying， microwave drying， freeze drying， sun drying， shade drying and combined drying）. The contents of 5 components such as gallic acid in wine-processed C. officinalis were determined by high- performance liquid chromatography. The contents of total flavonoids in wine-processed C. officinalis were determined by chromogenic method. Analytic hierarchy process was used to evaluate the effects of different drying methods on the contents of components in C. officinalis.RESULTS The contents of gallic acid， 5-hydroxymethylfurfural， monoside， loganin， cornuside and total flavonoids in 22 batches of wine-processed C. officinalis were 1.043 8-1.563 8， 0.648 5-2.358 8， 5.031 0-10.305 7， 6.681 2- 7.534 2， 0.986 5-1.148 8 and 33.657 2-50.741 5 mg/g， respectively. The comprehensive scoring results of analytic hierarchy process showed that the comprehensive score of each component in C. officinalis dried by microwave at 75 ℃ was higher ， followed by blast drying at 60 ℃ and far infrared drying at 60 ℃ .CONCLUSIONS The wine-processed C. officinalis could be dried by microwave drying at 75 ℃， blast drying at 60 ℃ or far infrared drying at 60 ℃.

METTL3 and METTL14 are two components that form the core heterodimer of the main RNA m6A methyltransferase complex (MTC) that installs m6A. Surprisingly, depletion of METTL3 or METTL14 displayed distinct effects on stemness maintenance of mouse embryonic stem cell (mESC). While comparable global hypo-methylation in RNA m6A was observed in Mettl3 or Mettl14 knockout mESCs, respectively. Mettl14 knockout led to a globally decreased nascent RNA synthesis, whereas Mettl3 depletion resulted in transcription upregulation, suggesting that METTL14 might possess an m6A-independent role in gene regulation. We found that METTL14 colocalizes with the repressive H3K27me3 modification. Mechanistically, METTL14, but not METTL3, binds H3K27me3 and recruits KDM6B to induce H3K27me3 demethylation independent of METTL3. Depletion of METTL14 thus led to a global increase in H3K27me3 level along with a global gene suppression. The effects of METTL14 on regulation of H3K27me3 is essential for the transition from self-renewal to differentiation of mESCs. This work reveals a regulatory mechanism on heterochromatin by METTL14 in a manner distinct from METTL3 and independently of m6A, and critically impacts transcriptional regulation, stemness maintenance, and differentiation of mESCs.
Animals ; Mice ; Methylation ; Chromatin ; Histones/metabolism* ; RNA, Messenger/genetics* ; Methyltransferases/metabolism* ; RNA/metabolism*