Objective:To establish the identification method for Aucklandia Lappa and Picrorhiza kurrooa Royle ex Benth and the content determination of hydroxysaffor yellow A in Liangxue Shiwei San. Methods:The identification was carried out by TLC. The con-tent of hydroxysaffor yellow A was determined by HPLC. The column was Kormasil C18 (250 mm × 4. 6 mm, 5 μm) and the flow rate was 1. 0 ml·min-1. The mobile phase was methanol-0. 5% acetic acid (30∶70). The detection wavelength was 403 nm and the col-umn temperature was 25℃,and the sample size was 10 μl. Results:The TLC spots were clear with high resolution without interference from the negative sample. A good linearity of hydroxysaffor yellow A was within the range of 1.212-48.480μg·ml-1(r=0.999 8). The average recovery was 98. 13%(RSD=1. 6%,n=6). Conclusion:The established TLC and HPLC methods are simple and accu-rate with good reproducibility, which can be used in the quality control of Liangxue Shiwei San.

Objective: To evaluate the effect of zinc ions on human umbilical vein endothelial cells biological functions. Methods: The primary human umbilical vein endothelial cells were cultured with the ECM medium, and cells were divided into 8 groups: the control group(routine culture,n=3), 20 μmol/L zinc group(20 μmol/L zinc chloride solution was added into the cell medium, n=3), 40 μmol/L zinc group(40 μmol/L zinc chloride solution was added into the cell medium, n=3),80 μmol/L zinc group(80 μmol/L zinc chloride solution was added into the cell medium, n=3), 100 μmol/L zinc group(100 μmol/L zinc chloride solution was added into the cell medium, n=3), 200 μmol/L zinc group(200 μmol/L zinc chloride solution was added into the cell medium, n=3),300 μmol/L zinc group(300 μmol/L zinc chloride solution was added into the cell medium, n=3), 500 μmol/L zinc group(500 μmol/L zinc chloride solution was added into the cell medium, n=3). The cell proliferation curve was derived from real time cell analysis (RTCA). The viability value was obtained via CCK-8 reagent, and the migration distance was tested by scratch-wound assay while the adhesion function was detected by RTCA. Results: (1)After 18 hours, RTCA showed that the proliferation cell indexes were 4.5±0.6, 3.7±0.4, 3.6±0.3, 2.5±0.4, and 2.5±0.4 in the 20, 40, 80, 100, and 200 μmol/L zinc groups, as compared with 3.5±0.3 in the control group (all P<0.05). Proliferation cell indexes were 0 in both of the 300 μmol/L and 500 μmol/L zinc groups. (2)After 96 hours, the viability were 1.21±0.05, 1.10±0.03, 0.99±0.05, 0.62±0.02, 0.45±0.04, 0.11±0.01, and 0.12±0.06, respectively in the 20, 40, 80, 100, 200, 300, and 500 μmol/L zinc groups, as compared with 0.75±0.05 in the control group (all P<0.05). (3)After 12 hours, the migration distances were (0.56±0.11),(0.96±0.07),(0.49±0.02), and (0.29±0.01)mm in the 20, 40, 80, and 100 μmol/L zinc groups, as compared with (0.24±0.04)mm in the control group (all P<0.05). (4)After 18 hours, the adhesion cell index were 0.40±0.05, 0.31±0.01, 0.38±0.05, and 0.40±0.03 in the 20, 40, 80, and 100 μmol/L zinc groups, as compared with 0.24±0.04 in the control group (all P>0.05). Conclusions Zinc ions at lower concentration (≤80 μmol/L) can promote proliferation, viability and migration of human umbilical vein endothelial cells, but the adhesion function was not significantly affected by zinc ions. Zinc ions at higher concentration (≥200 μmol/L) can inhibit the cellular function of the human umbilical vein endothelial cells.

Objective:To explore the therapeutic mechanism of Mongolian medicine Sendeng-4 decoction for rheumatoid arthritis by 99Tc m-hydrazinonicotinamide-(polyethylene glycol) 4-E[(polyethylene glycol) 4-c((Arg-Gly-Asp)fk)] 2 (3PRGD 2) imaging. Methods:A total of 200 female SD rats (age: 6-7 weeks) were divided into collagen-induced arthritis (CIA) group ( n=176) and blank control group ( n=24). Rats in the CIA group were divided into Sendeng-4 decoction treatment group ( n=24), etanercept treatment group ( n=24), and negative control group ( n=24) by simple random sampling method. 99Tc m-3PRGD 2 SPECT/CT imaging was performed before and after modeling and treatment. The differences of target/non-target (T/NT) ratio and serological, pathological, and immunohistochemical results among groups were compared by one-way analysis of variance or Kruskal-Wallis rank sum test. The correlation was analyzed by Pearson correlation or Spearman correlation analysis. Results:There were 95 (95/176) CIA models successfully established. The T/NT ratios of Sendeng-4 decoction treatment group and etanercept treatment group were lower than that of negative control group (0.260± 0.094, 0.238±0.099, 0.766±0.144 ; F=163.00, P<0.001), while there was no significant difference between the two drug treatment groups ( P>0.05). After drug treatment, serum levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-α (TNF-α) and α vβ 3 were significantly lower than those of negative control group ( F values: 49.43-92.36, all P<0.001), pathological score was also lower than that of negative control group ( H=34.25, P<0.001), and levels of immunohistochemical makers (VEGF, TNF-α, α vβ 3, CD31, CD34) were also lower than those of negative control group ( H values: 13.51-26.84, all P<0.001), while there were no significant differences between the two drug treatment groups (all P>0.05). The T/NT ratios were positively correlated with above indictors in Sendeng-4 decoction treatment group ( r values: 0.56-0.59, rs values: 0.49-0.69), etanercept treatment group ( r values: 0.50-0.55, rs values: 0.46-0.70) and negative control group ( r values: 0.55-0.80, rs values: 0.58-0.86, P<0.001 or P<0.05). Conclusion:Verified by 99Tc m-3PRGD 2 SPECT/CT imaging and molecular pathology, Mongolian medicine Sendeng-4 decoction can inhibit neovascularization by down-regulating vascular factors such as VEGF, resulting in delaying the progression of the disease and improving clinical symptoms.

Abstract OBJECTIVE To quickly qualitatively analyze the chemical components in Sanzi powder compound prescription by HPLC-Q-Exactive-MS/MS. METHODS SHIMADZU GIST C18 column(4.6 mm×150 mm, 5 μm) was used, using 0.1% formic acid water-methanol as mobile phase with gradient elution, the flow rate was 0.5 mL·min-1, the column temperature was 30 ℃. Under positive and negative ion modes, the primary and secondary mass spectrometry information of Sanzi powder was scanned. Qualitative attribution of each component in Sanzi powder was carried out based on the mass spectrometry information, molecular formula, and retention time of molecular ion peaks and fragment ions analyzed using total ion flow diagrams, combined with the molecular formula and structural formula searched in the Chemspider database and references. RESULTS Based on the analysis of the mass spectrometry cleavage rules and references of various components, 95 possible chemical components were preliminarily inferred, including 39 phenolic acids, 20 tannins, 9 organic acid esters, 5 monoterpenoids, 12 iridoids, 8 triterpenes and 2 flavonoids. Among them, 57 components were from Chebulae Fructus, 30 were from Gardeniae Fructus, 10 were from Toosendan Fructus, and among these compounds, rutin from Chebulae Fructus, Gardeniae Fructus and Toosendan Fructus. CONCLUSION The HPLC-Q-Exactive-MS/MS detection method has good separation and high sensitivity, and can quickly and efficiently infer various components in Sanzi powder. It establishes a fast and efficient analytical method for identifying the chemical components in Sanzi powder.

OBJECTIVE To establi sh the method for the con tent determination of 11 components in Terminalia chebula from different origins ，and to provide reference for their quality evaluation and superior provenance screening. METHODS Taking 16 batches of T. chebula from different origins as test samples ，high performance liquid chromatography tandem triple quadrupole mass spectrometry was established to determine the contents of 11 components，such as vitexin ，gallic acid ，methyl gallate ，ethyl gallate，ellagic acid ，corilagin，shikimic acid ，ferulic acid ，luteolin，quercetin and rutin. The determination was performed on Shim-pack GIST-HP C 18 column with mobile phase consisted of 0.1％ formic acid solution-methanol at the flow rate of 0.25 mL/ min（gradient elution ）. The sample size was 3 μL，and the column temperature was 35 ℃. Electrospray ionization source was used in positive and negative ion mode ，with multiple reaction monitoring. The atomized gas flow rate was 3 L/min，the heating gas flow rate was 10 L/min，the interface temperature was 300 ℃，the desolvent temperature was 526 ℃，and the heating block temperature was 400 ℃ . Grey correlation analysis （GRA）and technique for order preference by similarity to ideal solution （TOPSIS）methods were used to compare ，analyze and comprehensively evaluate T. chebula from different origins. RESULTS The results of content determination methodology met the relevant requirements. The contents of 11 components in 16 batches of T. chebula were 7.27-106.38，5 370.24-31 010.43，21.42-1 097.50，4.26-111.09，17 940.42-38 490.18，6 247.26-40 182.18，12 125.94- 209 519.96，2.71-9.04，0.24-44.12，1.49-9.17 and 25.35-126.51 μg/g，respectively. The results of GRA and TOPSIS analysis showed that the comprehensive qualities of sample H 12（from Yunnan ），H11（from Guangxi ），H5（from Hunan ），H14（from Guangdong），H13（from Sichuan ），H8（from Guangdong ），H1（from Yunnan ）were better. CONCLUSIONS The established method is fast ，sensitive and reliable ，and can be suitable for comprehensive evaluation of the internal quality and superior provenance screening of T. chebula .

OBJECTIVE To establish the HPLC fingerprint of Mongolian medicine Sanzisan ，and to evaluate its internal quality by chemical pattern recognition technique comprehensively. METHODS HPLC method was used. Using geniposide as reference，HPLC fingerprints of 15 batches of Sanzisan were drawn with Similarity Evaluation System of TCM Chromatogram Fingerprint（2012 edition）. Similarity evaluation and common peaks identification were conducted. Combined with cluster analysis （CA），principal component analysis （PCA），and orthogonal partial least squares-discriminant analysis （OPLS-DA），the quality of 15 batches of Sanzisan was evaluated ，and the differential markers that affected its quality were screened. RESULTS There were 29 common peaks in 15 batches of Sanzisan ，and the similarity was no less than 0.952，indicating that the chemical composition of the 15 batches of Sanzisan had good consistency. A total of 13 common peaks were identified ，which were chebulic acid ，gallic acid，punicalin，punicalagin A ，punicalagin B ，jasminoside B ，caffeic acid ，corilagin，geniposide，chebulagic acid ，1，2，3，4，6- O-galloylglucose，chebulinic acid ，ellagic acid. Both CA and PCA could divide 15 batches of Sanzisan into four categories ，and the classification results were consistent ，indicating that the quality of 15 batches of Sanzisan had certain differences. Fourteen differential markers （chebulic acid ，gallic acid ，ellagic acid ，etc）that lead to the quality difference between batches were screened out by OPLS-DA. CONCLUSIONS Established HPLC fingerprint analysis method is simple and stable. Combined with chemical pattern recognition analysis ，it can be used for the quality control of Sanzisan.

OBJECTIVE To establish the methods to identify the chemical components of Ixeris chinensis， and determine the contents of 7 components （chlorogenic acid， luteolin， quercetin， rutin， protocatechuic acid， isochlorogenic acid A， luteoloside）. METHODS HPLC-Q-Exactive-MS was used to identify the chemical components of I. chinensis. The contents of 7 components in I. chinensis， including chlorogenic acid， were determined by HPLC-MS/MS. RESULTS A total of 45 components were identified in I. chinensis， including 20 organic acids， 13 flavonoids， 4 fatty acids， 4 amino acids， 3 nucleosides， and 1 coumarin. The linear range of chlorogenic acid， luteolin， quercetin， rutin， protocatechuic acid， isochlorogenic acid A and luteoloside were 503.00- 25 150.00， 42.00-2 100.00， 5.05-252.50， 20.05-1 002.50， 25.10-1 255.00， 750.00-37 500.00， 196.00-9 800.00 ng/mL （r≥0.999 2）， respectively. RSDs of precision， stability and reproducibility tests were all less than 3.00% （n＝6）， and average recovery ranged from 96.72% to 105.84% （all RSD＜4.00%， n＝6）. The contents of 7 components in 3 batches of I. chinensis were 1 145.77- 3 261.25， 23.75-97.90， 0.92-2.12， 1.06-23.18， 9.35-21.85， 833.25-1 045.58， 199.56-1 869.78 μg/g， respectively. CONCLUSIONS The established methods for identification and content determination are rapid and simple， and can be used for the identification of chemical components and the content determination of 7 components in I. chinensis.