OBJECTIVE To optimize the processing technology of Epimedium koreanum roasted with suet oil and analyze its characteristic chromatogram before and after processing. METHODS The optimal processing technology was optimized by orthogonal experiments with frying power， frying time and medicinal temperature as factors using the comprehensive score of appearance traits （color+odor）， alcohol-soluble extract， the contents of icariin and baohuoside Ⅰ as evaluation index， then proceed with verification. The E. koreanum roasted with suet oil was prepared with the optimal technology. The characteristic chromatograms of 15 batches of E. koreanum were established with the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition）， and then similarity analysis was also conducted. Clustering analysis， principal component analysis， and orthogonal partial least squares discriminant analysis were used to evaluate the differences in E. koreanum before and after processing. RESULTS The optimal processing technique for E. koreanum roasted with suet oil was as follows： first， heat 4 g of suet oil at a power of 600 W until it melts； next， when the temperature at the bottom of the pan reaches 140 ℃， add 20 g of E. koreanum silk and stir-fry for 6 minutes； finally， remove it and let it cool. Comprehensive score of 3 validation tests was 98.94 points　（RSD＜3%）. The established characteristic chromatogram of E. koreanum and E. koreanum roasted with suet oil were calibrated with 16 and 15 common peaks， respectively. Chromatographic peak 2 was determined to be chlorogenic acid， peak 5 to be chaohuoding A， peak 6 to be chaohuoding B， peak 7 to be chaohuoding C， peak 8 to be icariin， and peak 14 to be baohuoside Ⅰ. The similarities were all greater than 0.9. Results of cluster analysis and principal component analysis showed that E. koreanum and E. koreanum roasted with suet oil were clustered into distinct groups. The results of orthogonal partial least squares discriminant analysis showed that the variable importance projection values for peak 13， peak 15， peak 9， peak 1， peak 8， peak 12， peak 7， and peak 10 were all greater than 1. CONCLUSIONS The study successfully optimized the processing technology of suet oil-roasted E. koreanum. There are significant differences in the characteristic chromatograms of E. koreanum before and after processing. Among them， the chemical components corresponding to peak 13， peak 15， peak 9， peak 1， peak 8， peak 12， peak 7， and peak 10 may be the differential components of E. koreanum before and after processing.

Objective:To screen the differentially expressed genes(DEGs)under high phosphate-induced calcification in the vascular smooth muscle cells(VSMCs)by mRNA high-throughput sequencing technology,and to analyze the key genes and signaling pathways involved in the VSMCs calcification.Methods:The human VSMCs were divided into control group and model group.The cells in model group was exposed to the high-phosphate medium,while the cells in control group were cultured in DMEM supplemented with 10%fetal bovine serum under the same conditions.The VSMCs in two groups,stably transfected,were cultured for 12 d.The morphology of the cells in two groups were observed and photographed under inverted microscope.The DEGs were selected by Hisat2 software,and Gene Ontology(GO)functional and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were performed by Stringtie software from three aspects,such as biological processes(BP),molecular functions(MF),and cellular components(CC).The calcification of the cells in two groups was observed by Von Kossa staining method.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to analyze the expression levels of alkaline phosphatase(ALP),bone morphogenetic protein 2(BMP2),alpha-smooth muscle actin(α-SMA),tumor protein 53(Tp53),glutathione peroxidase 4(GPX4),ferritin light chain 1(Ftl1),and glycosylphosphatidylinositol-specific phospholipase D1(GPLD1)mRNA in the cells in two groups.Results:Compared with control group,there were 2 524 DEGs in the cells in model group,and there were 1 368 upregulated DEGs and 1 156 downregulated DEGs.Clustering of DEGs between the cells in two groups was distinct.The GO functional and KEGG pathway enrichment analysis results showed that the upregulated DEGs were primarily involved in regulating the microtubule cytoskeleton,cell polarity,protein localization,and cell cycle regulation among BPs;in constructing cell membrane,microtubule organization,chromosomes,and kinetochore among CCs;and functioning in phosphatidylinositol phosphate,Rho GTPase protein binding,transmembrane transport,and protein kinase regulatory activity among MFs.Downregulated DEGs were mainly involved in cytoplasmic translation,protein membrane localization,mRNA metabolism,and protein endoplasmic reticulum localization among BPs;in forming ribosome subunits,cell membrane,and autophagy among CCs;and functioning in single-stranded DNA,ribonucleoprotein complex,growth factor binding,regulating protein kinase activity,and catalytic activity among MFs.Seven signaling pathways were significantly enriched in upregulated genes,most notably in the biosynthesis of glycosylphosphatidylinositol(GPI)anchors;whereas 18 signaling pathways were significantly enriched in the downregulated genes,most notably in ferroptosis.The RT-qPCR results showed that compared with control group,the expression levels of GPX4,Ftl1,and Tp53 mRNA in the cells in model group were significantly decreased(P<0.01),while the expression level of GPLD1 mRNA was significantly increased(P<0.01);compared with control group,the expression level of α-SMA mRNA in the cells in model group was significantly decreased(P<0.01),and the expression levels of ALP and BMP2 mRNA were significantly increased(P<0.01).Conclusion:The VSMCs underwent calcification and normal cells exhibit the DEGs.The key signaling pathways in the calcification induced by high phosphate in the VSMCs include ferroptosis and GPI anchor biosynthesis,mediated primarily through GPX4,Ftl1,Tp53,and GPLD1.

OBJECTIVE To optimize the preparation process of Soft-shelled turtle blood lyophilized powder （STBLP）， and to provide a reference for improving the availability and quality stability of soft-shelled turtle blood （STB）. METHODS STBLP was prepared with vacuum freeze-drying. Taking the solubility as the index， the preparation process parameters of STBLP were optimized by single factor experiment and Box-Behnken response surface method. RESULTS The optimal freeze-drying process for STBLP was obtained： pre-freezing time of 4 h， total drying time of 13 h （before at 0 ℃）， and resolution drying temperature of 25 ℃. The average solubility of 3 batches of STBLP prepared according to the optimal process was 95.72% （RSD＝0.68%， n＝3）， the relative error of which was －0.97% to the theoretical solubility （96.66%）. CONCLUSIONS Optimized lyophilization process in this study are stable and feasible， the solubility of the prepared sample is high.

OBJECTIVE To analyze the odor composition changes of two kinds of traditional Chinese medicine sachet （children type and adults type） with different placement time by using ultra-fast gasphase electronic nose technology. METHODS The change rule of sachet components at different storage times was analyzed by gas chromatography. At the same time， the qualitative results were obtained by combining electronic nose with Arochembase database. Discriminant factor analysis was used to analyze the overall odor composition differences of the two sachet samples. RESULTS A total of 10 odor compositions were identified in children-type sachet， including α-pinene and β-pinene as the functional index compositions； five odor compositions of children-type sachet disappeared after 0.25 days， and most of them disappeared after 7 days； the cumulative contribution rate of discriminant factor analysis was 99.225%. A total of 8 odor compositions were identified in adult-type sachets， including α-pinene and α-phellandrene as the functional index compositions； four odor components disappeared after the adult-type sachet was placed for 0.25 days； after 15 days of placement， the peak 6-8 disappeared， and the intensity of peak 5 decreased by 34.3% compared with 0 day of placement； the cumulative contribution rate of discriminant factor analysis was 91.965%. CONCLUSIONS With the extension of storage time， the smell and composition of the two traditional Chinese medicine sachets are decreasing. It is recommended that the use time of children-type sachet is 7 days， and that of adult-type sachet is 15 days.

OBJECTIVE To establish the method for monitoring the dynamic changes of odor components in Cornus officinalis during processing . METHODS The decoction pieces of C. officinalis with different processing time were prepared by the wine steaming method . The dynamic changes of odor components were obtained by using ultra -fast gas electronic nose ；odor components were identified by comparing with AroChemBase database ；the dynamic changes of odor compounds were analyzed in combination with peak area ，and the chemical pattern recognition analysis were carried out . RESULTS A total of 12 common peaks of odor components were identified in the fingerprints of raw C. officinalis，and 21 in the fingerprints of decoction pieces of C. officinalis. Eight odor components with the high proportion of peak area during processing were ethanol ， isopropyl alcohol ， 2- methylpropylaldehyde，ethyl acetate ，2-methylbutanal，isoamyl alcohol ，2-hexanol and furfural ，among which ，the peak areas of ethanol，isoamyl alcohol and 2-hexanol showed a trend of first increasing and then decreasing ；at 24 h of processing ，their peak areas were still higher than those of raw products . The peak areas of ethyl acetate ，2-methylbutanal and furfural nearly increased with the increase of processing time . Variable importance in projection of above eight odor components were all greater than 1. CONCLUSIONS The method is established for monitoring the dynamic changes of odor components of C. officinalis during processing. Eight odor components such as ethanol can be used as monitoring indicators of C. officinalis dring processing .

ObjectiveIn order to find a fast odor-based method for the identification of sulfur fumigated Gastrodiae Rhizoma， an ultra-fast gas phase electronic nose technology was used to identify the odors of different degrees of sulfur fumigated Gastrodiae Rhizoma decoction pieces. MethodHeracles NEO ultra-fast gas phase electronic nose was employed to collect gas chromatograms of unsulfured and sulfured with different degrees of Gastrodiae Rhizoma decoction pieces， gas chromatograms were performed under programmed temperature （initial temperature of 40 ℃， 0.2 ℃·s^-1 to 60 ℃， and then 4 ℃·s^-1 to 250 ℃）， the sample volume was 5 mL， the incubation temperature was 65 ℃ and incubation time was 35 min. Kovats retention index and the AroChemBase database were used for qualitative analysis， and stoichiometric analysis was performed on this basis. Principal component analysis （PCA）， discriminant factor analysis （DFA） and partial least squares-discriminant analysis （PLS-DA） models were established to identify the Gastrodiae Rhizoma decoction pieces with different degrees of sulfur fumigation. ResultAccording to the comparative analysis of AroChemBase database， there were significant differences in the odor characteristics of sulfur fumigated and non-sulfur fumigated Gastrodiae Rhizoma， cyclopentane， acetone and heptane might be the odor components to distinguish the degree of sulfur fumigation in Gastrodiae Rhizoma decoction pieces. The identification index of PCA model was 81， the accumulative discriminant index of the discriminating factors was 92.09% in DFA model， the supervisory model interpretation rate of PLS-DA model was 0.963 and the predictive ability parameter was 0.956， indicating that PCA， DFA and PLS-DA models could well distinguish Gastrodiae Rhizoma decoction pieces with different sulfur fumigation degrees. ConclusionHeracles NEO ultra-fast gas phase electronic nose can be used as a rapid method to identify and distinguish Gastrodiae Rhizoma decoction pieces with different levels of sulfur fumigation. Meanwhile， it can provide a rapid， simple and green method and technology for identification of traditional Chinese medicine decoction pieces by sulfur fumigation.

OBJECTIVE To establish the fingerprints of dried Houttuynia cordata and its decoction pieces ，conduct chemometrics analysis and determine the contents of 5 flavonoids such as neochlorogenic acid. METHODS High performance liquid chromatography （HPLC）method was adopted. Using quercitrin as reference ，HPLC fingerprints of 10 batches of dried H. cordata and its decoction pieces were drawn. The similarity evaluation was conducted by Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition），the common peaks were also confirmed. SIMCA-P 14.1 software was applied for principal component analysis （PCA）and partial least square-discriminant analysis （PLS-DA），and the variable importance in projection（VIP）value more than 1 was considered as a standard to screen the differential components affecting the quality of these two products ；meanwhile，the contents of 5 components such as neochlorogenic acid in both products were determined by the same HPLC method. RESULTS There were 20 common peaks in 10 batches of dried H. cordata and 10 batches of its decoction pieces with the similarity values more than 0.960. A total of 5 common peaks were identified ，which were neochlorogenic acid （peak 1）， chlorogenic acid （peak 3），cryptochlorogenic acid （peak 4），rutin（peak 7）and quercitrin （peak 11）. The results of PCA and PLS-DA showed that dried H. cordata could be distinguished from its decoction pieces obviously ；the common peaks with VIP value greater than 1 were as follows ：peak 7（rutin），peak 20，peak 5，peak 13，peak 2，peak 18，peak 3（chlorogenic acid ）， peak 14，peak 17 and peak 19. The linear range of neochlorogenic acid ，chlorogenic acid ，cryptochlorogenic acid ，rutin and quercitrin were 3.77-60.29 μg/mL（r＝0.999 7），1.40-22.42 μg/mL（r＝0.999 5），3.76-60.22 μg/mL（r＝0.999 9），2.19-35.06 μg/mL （r＝0.999 9）and 25.49-407.88 μg/mL（r＝0.999 7），respectively. RSDs of precision ，stability（24 h）and reproducibility E-mail：20190394@njucm.edu.cn tests were all lower than 3％. The average recoveries of the above components in these two products were 98.72％-101.12％ and 98.86％ -100.63％ with RSDs less than 3％（n＝9）. In dried H. cordata ，the average contents of 5 components were 0.87，0.33，0.59，0.61 and 6.17 mg/g，while the average contents were 0.42，0.11，0.26，0.23 and 3.16 mg/g in its decoction pieces ，respectively. CONCLUSIONS HPLC fingerprint and the method of content determination are stable and feasible ，which could be used for the quality control of dried H. cordata and its decoction pieces. Besides ，rutin and other components may be the differential components which could affect the quality of these two products ；the average contents of the 5 flavonoids such as neochlorogenic acid in dried H. cordata all decrease after processing.

OBJECTIVE：To esta blish the m ethod for identifying Schizonepeta tenusfolia from different habitats based on odor information. METHODS ：The odor of S. tenusfolia from different habitats were identified by Heracles Ⅱ ultra-fast gas phase electronic nose. Qualitative analysis was conducted according to obtained chromatographic information combined with AroChemBase database and Kovats retention index qualitative database. Principle component analysis （PCA）and discriminant factor analysis （DFA）were conducted by using Alpha Soft V 14.2 software，and cluster analysis （CA）was performed with SPSS 22.2 software. RESULTS ：There were 16 common peaks in 15 batches of S. tenusfolia from different habitats. After comparison with AroChemBase database and Kovates retention index qualitative database ，a total of 13 possible components were obtained. The possible components and sensory description information of S. tenusfolia from different habitats were basically the same ，but only the content was different. The chromatographic peak intensities of common peak No. 2 were in descending order as Anhui ＞ Gansu＞Henan＞Hebei＞Jiangsu，the chromatographic peak intensities of common peak No. 6 were in descending order as Anhui ＞ Hebei＞Gansu≈Henan＞Jiangsu，the chromatographic peak intensities of common peak No. 9 were in descending order as Anhui ＞ Gansu＞Henan＞Jiangsu＞Hebei，the chromatographic peak intensity of common peak No. 13 were in descending order as Anhui ≈ Gansu＞Hebei＞Jiangsu＞Henan，which represented the chromatographic peak intensity of methyl formate （peak No. 2），α-pinene （peak No. 6），3-nonone（peak No. 9）and α-terpineol（peak No. 13）were significantly different due to the change of habitats. PCA results showed that the cumulative contribution rate of the first two principal components was 96.807％. Results of DFA showed that contribution rates of discriminant factor 1 and discriminant factor 2 were 92.089％ and 3.982％. CA results showed that when the distance was 10，15 batches of samples could be clustered into 3 categories，B1-B5 and J 1-J3 into one category ，A1-A3 into one category ，G1，G2，N1 and N 2 into one category. The results were basically consistent with those of PCA and DFA. CONCLUSIONS：Ultra-fast gas phase electronic nose technology can be used to identify S. tenusfolia from different habitats rapidly. Methyl formate ，α-pinene，3-nonone and α-terpineol may be the key factors to distinguish S. tenusfolia from different habitats.

AIM To develop an HPLC method for the simultaneous content determination of five constituents in Chairen Capsules (Bupleuri Radix,Ziziphi spinosae Semen,Chuanxiong Rhizoma,etc.).METHODS The analysis of 80％ methanol extract of this drug was performed on a 30 ℃ thermostatic Merck Purospher STAR C1s column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ formic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 237,290 and 335 nm.RESULTS Spinosin,ferulic acid,cinnamaldehyde,liquiritin and ammonium glycyrrhizate showed good linear relationships within the ranges of 0.009 26-0.092 6 μg (R2 =0.999 6),0.056 2-0.562 μg (R2 =0.999 8),0.130-1.30 μg (R2 =0.999 8),0.275-2.75 μg (R2 =0.999 5) and 0.568-5.68 μg (R2 =0.999 2),whose average recoveries were 100.73％,100.62％,100.02％,100.41％ and 99.96％ ％ with the RSDs of 2.20％,1.37％,1.52％,1.30％ and 0.80％,respectively.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Chairen Capsules.

OBJECTIVE:To establish a method for the simultaneous determination of rutin ,hesperidin ,arctiin ,liquiritin and glycyrrhizinate in Sangxing oral liquid. METHODS:RP-HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.1% phosphoric acid solution(gradient elution) at a flow rate of 1.0 mL/min,the detection wavelength was 276, 348 nm ,column temperature was 30 ℃,and the injection volume was 5 μL. RESULTS:The linear range was 1.068-10.68 μg/mL (r=0.9993)for rutin,9.320-93.20 μg/mL(r=0.9992)for hesperidin,46.91-469.1 μg/mL(r=0.9994)for arctiin,0.5110-5.110μg/mL(r=0.9992) for liquiritin;RSDs of precision , stability and reproducibility tests were less than 3%;recoveries were 95.10% -101.90%(RSD=2.1%,n=9),96.75%-101.80%(RSD=1.4%,n=9),95.69%-100.00%(RSD=1.5%,n=9), 98.22%-104.60%(RSD=2.1%,n=9),respectively. CONCLUSIONS:The method is accurate,sensitive and reproducible,and can be used for the simultaneous determination of rutin,hesperidin,arctiin and liquiritin in Sangxing oral liquid.