ABSTRACT
OBJECTIVE：To establish a method for simultaneous determination of the contents of codonopatin ，syringin， atractylenolide Ⅰ，atractylenolide Ⅱ and atractylenolide Ⅲ，and to compare the contents of above 5 components in different varieties and harvesting periods of Codonopsis Radix. METHODS ：HPLC method was used. The column was Inertsil ODS- 3 with mobile phase consisted of acetonitrile-water （gradient elution ）at the flow rate of 0.8 mL/min. The detection wavelengths were 210 nm （codonopatin），220 nm （syringin，atractylenolide Ⅱ ，atractylenolide Ⅲ），276 nm （atractylenolide Ⅰ）. The column temperature was set at 30 ℃ ，and the sample size was 20 μ L. RESULTS：The linear range of codonopatin ，syringin， atractylenolide Ⅰ，atractylenolide Ⅱ and atractylenolide Ⅲ were 44.30-886.00 μg/mL（r＝0.999 7），6.50-130.03 μg/mL（r＝0.999 6）， 4.47-89.46 μg/mL（r＝0.999 5），2.53-50.50 μg/mL（r＝0.999 4），5.64-112.80 μg/mL（r＝0.999 5）；the limits of quantification were 2.446 0，0.168 0，0.248 1，0.065 7，0.099 8 μg/mL，and detection limits were 1.352 0，0.067 2，0.005 4，0.006 3，0.007 3 μ g/mL；RSDs of precision ，stability（24 h），repeatability and durability tests were all less than 2％；the recoveries were 98.87％-100.62％（RSD＝0.73％，n＝6），98.46％-101.54％ （RSD＝1.15％，n＝6），98.32％-101.12％（RSD＝1.19％，n＝ 96.83％-104.16％（RSD＝2.62％，n＝6），97.87％-100.99％ （RSD＝1.07％，n＝6）. The average contents were 33.78-431.82， 0-20.60，0.44-3.68，0-10.83，0.27-73.40 μ g/g. The content of 1271985629@qq.com codonopatin was in descending order was as follows as Codonopsis pilosula ＞C. tangshen ＞C. pilosula Nannf. var. modesta （Nannf.） L. T. Shen ＞ecotypic variety of C.·1677· tangshen. The content of syringin in descending order was C. pilosula ＞C. pilosula Nannf. var. modesta（Nannf.）L. T. Shen ＞C. tangshen，but it was not detected in ecotypic variety of C. tangshen . The content of atractylenolide Ⅰ in descending order was C. pilosula Nannf. var. modesta（Namf.）L. T. Shen ＞ecotypic variety of C. tangshen ＞C. pilosula ＞C. tangshen . The content of atractylenolide Ⅱ in C. pilosula was higher than C. pilosula Nannf. var. modesta（Nannf.）L. T. Shen ，but was no detected in C. tangshen and ecotypic variety of C. tangshen . The content of atractylenolide Ⅲ in descending order was C. pilosula ＞C. pilosula Nannf. var. modesta（Nannf.）L. T. Shen ＞ecotypic variety of C. tangshen ＞C. tangshen . In Codonopsis Radix collected from Jul. to Oct. ，the content of codonopatin was the highest ；the content of atractylenolide Ⅰ was lower in sample collected from Jun. to Oct.；atractylenolide Ⅱ was not detected in sample collected in Aug. ；the contents of atractylenolide Ⅰ and atractylenolide Ⅱ were the lower in sample collected in Sept. ，and syringin and atractylenolide Ⅱ were not detected in some samples. CONCLUSIONS ： The established HPLC method is simple ，accurate，highly sensitive and reproducible. It can be used to simultaneously determine 5 active ingredients contents of Codonopsis Radix ；there are great difference in contents of 5 active ingredients in different varieties and harvesting periods of Codonopsis Radix.
ABSTRACT
Objective To investigate the rules of root tuberenlargement and medicinal materials formation, and lay the foundation for breeding a new Ophiopogon japonicus variety. Method The study was carried out with field cultivation. Samples were collected continuously for every interval 7-10 d. The rules of dry matter distribution and accumulation in each parts of O. japonicus was calculated based on the mean value of 15 plant samples. The number and size of root tuber were calculated based on the mean value of 15 samples. The fiber characteristics of root tuber of O. japonicus were observed by optical microscope. Characters of O. japonicus were observed according to the Chinese Pharmacopoeia 2015 edition and recorded the changes of medicinal characteristics. Results The root tuber of O. japonicus inflated starting from October to April in the next year in Sichuan province. Adventitious roots were initially formed into nutritive roots around caudex, and able to form into root tuber around caudex later. Tuber number of per plant is basic from 5 to 20, most in 8-10, very few more than 20. Adventitious roots expanded gradually into a fusiform root from one end of root tip, root tip can continue to grow to form adventitious roots after the formation of root tuber, the newly formed adventitious roots could be expanded for the second time, very few might appear swollen three times to form beads. Microscopic observation showed that root tubers were inflated primarily by the enlargement of cortical cells and the increase of cell layers, in which cortical cells enlargement played a leading role. While tubers started expanding, the accumulation of dry matter in fibrous roots and stem leaves quickly slowed to stop eventually, and the accumulation of root dry matter increased rapidly in the early stage, and it kept basically dynamic balance in mid and late March, and the whole behavior of medicinal materials was in the process from dry to full. Conclusion The results show the apparent change rules of O. japonicus root tuber in the expanding process, which proves the direction and goal for research on the mechanism of root tuber development of O. japonicus.
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Objective To evaluate the role of miR219‐2‐3p in gastric cancer and the underlying mechanisms .Methods Real time RT‐PCR was employed to quantify the expression level of miR219‐2‐3p in matched tissues of 82 patients .In vitro cell proliferation , the expression of tumorgenesis related protein ERK1/2 was performed by overexpressing miR‐219‐2‐3p in gastric cancer cell line MGC803 .Results The expression level of miR‐219‐2‐3p significantly decreased in late stage gastric cancer (P< 0 .05) .The cell proliferation rate was significantly reduced in gastric cancer cell lines overexpressing miR‐219‐2‐3p .Furthermore ,the expression level of p‐ERK that activated form of ERK was significantly decreased whereas the total level of ERK was unchanged after overex‐pressing of miR‐219‐2‐3p in MGC803 cell lines .The expression level of p‐ERK was also increased in gastric cancer tissues .Conclu‐sion miR‐219‐2‐3p may acts as a tumor suppressor in gastric cancer by decreasing the activity of ERK 1/2 signaling pathway .