OBJECTIVE To identify Ginseng Flos and their confounds by using the high-throughput sequencing technology, and to verify the accuracy of high-throughput sequencing technology in species identification by using ITS2 sequencing technology. METHODS High-throughput sequencing was performed on the amplified products of Ginseng Flos adulterated samples, use cutadapt, PEAR, PRINSEQ, Usearch, RDP classifier, SINTAX software to obtain operational taxonomic unit(OUT) sequences, remove fungi, unclassified and other non-green plant sequences. To avoid false positives, delete OTU sequences with a sequence number <100 or base numbers <200 bp. The ITS2 amplification products of Ginseng Flos, Quinquefolii Flos, and Notoginseng Flos were sequenced. To verify the accuracy of high-throughput sequencing technology for species identification, MEGA 11.0 was used to construct neighbor joining system cluster tree, genetic distance, interspecific information loci and Blast analysis of ITS2 and OTU base sequences of Ginseng Flos, Quinquefolii Flos, and Notoginseng Flos. RESULTS A total of 54 653 valid sequences were obtained by high-throughput sequencing, the serial numbers of Ginseng Flos, Quinquefolii Flos, and Notoginseng Flos were OTU1, OTU2, OTU3, respectively, and the corresponding effective sequences were 31 325, 857 and 442, respectively. By performing a Blast search of ITS2 and OTU base sequences of each species, each species was supported. The genetic distance between Ginseng Flos and Quinquefolii Flos and Notoginseng Flos was 0.010 and 0.033, respectively. Ginseng Flos and Quinquefolii Flos, Notoginseng Flos had 2 and 7 information sites, respectively. The neighbor join system cluster tree showed that the species were clustered independently into one branch, with Ginseng Flos, and Quinquefolii Flos clustered as a large branch and juxtaposed with Notoginseng Flos. Ginseng Flos was the same as Quinquefolii Flos secondary structure, but with Notoginseng Flos there were three different positions but there were A, B and C differences between arm Ⅳ and arm Ⅰ of Notoginseng Flos. CONCLUSION The high-throughput sequencing technology can accurately identify Ginseng Flos, Quinquefolii Flos and Notoginseng Flos, and has a strong ability to identify adulterated samples, which provides a certain idea for the identification of commercial Ginseng Flos.

OBJECTIVE：To esta blish a method for si multaneous determination of 5 components in the branch and root of Juglans mandshurica as gallic acid ，ellagic acid ，1，6-di-O-galloyl-β-D-glucose，1，2，6-tri-O-galloyl-β-D-glucose and 1，2，3， 6-tetra-O-galloyl-β-D-glucose，and to analyze the content difference of above 5 components between the branch and root samples. METHODS：HPLC method was adopted. The determination was performed on Agilent Poroshell 120 SB-C18 column with mobile phase consisted of water （containing 0.2％ formic acid ）-acetonitrile （containing 0.2％ formic acid ）. A gradient elution was performed at a flow rate of 0.3 mL/min. The column temperature was 30 ℃ and the detection wavelength was 270 nm. The sample size was 5 μL. Independent samples t-test and partial least squares-discriminant analysis （PLS-DA）were applied for statistical analysis of 5 components. RESULTS ：The linear range of gallic acid ，ellagic acid ，1，6-di-O-galloyl-β-D-glucose，1，2， 6-tri-O-galloyl-β-D-glucose and 1，2，3，6-tetra-O-galloyl-β-D-glucose were 0.989-63.3，1.58-101，1.01-64.7，3.31-212，3.34-214 μg/mL （r≥0.997 3），respectively. RSDs of precision ，reproducibility and stability tests （12 h）were all lower than 3.2％. The average recoveries of the 5 components were 103.2％（RSD＝4.85％），99.1％（RSD＝2.80％），101.5％（RSD＝1.31％），102.9％（RSD＝ 2.73％）and 104.7％（RSD＝1.28％），respectively. The average contents of the above components in the branch of J. mandshurica were 0.296 5，0.621 1，0.562 5，3.111 7 and 3.451 3 mg/g，respectively. The average contents of above components in the root were 0.673 4，2.755 5，0.964 0，2.946 6 and 4.836 4 mg/g，respectively. The total contents of the 5 components in the branch and roo t of J. mandshurica were 8.043 2 and 12.175 9 mg/g，respectively. The contents of gallic acid ，ellagic acid and 1，6-di-O-galloyl- β-D-glucose in roots were significantly higher than those in branches （P＜0.05 or P＜0.01）. There were no significant differences in the contents of the other 2 components and the total contents of the 5 components in branches and roots （P＞0.05）. The cumulative interpretability （R 2X，R 2Y） and cumulative predictability （Q 2） of the model established by PLS-DA were 0.943，0.745，and 0.710 respectively. The model load diagram showed that the distance between the ellagic acid and the origin was the farthest ，and only variable projection importance of the content of the ellagic acid was greater than 1. CONCLUSIONS：The established method can be used for the content determination of 5 components in the branch and root of J. mandshurica . Except for 1，2，6-tri-O-galloyl-β-D-glucose，the contents of other 4 components and total contents of the 5 components in the root of J. mandshurica are higher than those of the branch. Ellagic acid is selected as the potential marker for discriminating the branch and root samples.

OBJECTIVE：To investigate the internal mechanism of Schisandra sphenanthera and Schisandra chinensis in determining quality by color （“color discrimination grading ”）of medicinal materials ，and to construct a qualitative identification model based on color quantization value. METHODS ：HPLC method was used to determine the contents of 6 active components from 39 batches of samples. The colorimeter was used to determine 3-color spatial value [lightness value （ΔL＊），red-green value （Δa＊），yellow-blue value （Δb＊）]. SPSS 24.0 statistical software was used to analyze the correlation between the contents of 6 active components and 3-color spatial values. Principal component analysis （PCA）was performed by using SIMCA-P 14.1 software. RESULTS：The linear range of schizandrol A ，schizandrol B ，schisandrin A ，schisandrin B ，schisandrin C ，schisantherin A were 0.204 8-2.560 0，0.049 3-0.616 3，0.098 4- 1.230 0，0.046 3-0.578 8，0.010 6-0.132 0，0.100 0-1.500 0 μg（r＞0.999 0）；RSDs of precision ，stability（12 h）and repeatability tests were all less than 3％. The recoveries were 98.14％-101.53％（RSD＝1.08％， n＝6），97.16％-101.05％（RSD＝1.54％，n＝6），98.29％-101.41％（RSD＝1.29％，n＝6），97.17％-100.36％（RSD＝1.20％，n＝ 6），97.32％-102.43％（RSD＝1.77％，n＝6）and 98.02％-100.40％（RSD＝0.84％，n＝6），respectively. Among 39 batches of components were 3.25-7.39，0.96-1.98，0.46-4.74，1.62-2.60， 0.06-0.58，0.48-6.11 mg/g，respectively. Average S. chinensis was － 80.79-－ 70.54， average Δ a ＊ was qq.com # 通 2.54-5.34，average Δb＊ was 5.20-12.83，average ΔE＊ was 71.13-81.23；average ΔL＊ of S. sphe nanthera was －75.90- －69.16，average Δa＊ was 3.77-7.82，average Δb＊ was 8.59-17.23，average ΔE＊ was 69.99-77.92. The results of relationship analysis showed that the contents of schizandrol A ，schizandrol B ，schisandrin A ，schisandrin B and schisantherin A were significantly correlated with ΔL＊，Δa＊，ΔE＊（P＜0.01），with no significant correlation with Δb＊（P＞0.05）. There was a negative correlation of the content of schisandrin C with ΔL＊ and Δa＊（P＜0.05），and there was no significant correlation with Δb＊ and ΔE＊ （P＞0.05）. Results of PCA showed that accumulative variance contribution rate of primary 2 main components was 89.8％，and S. sphenanthera and S. chinensis could be identified significantly. CONCLUSIONS ：The content of schizandrol A in S. chinensis is high relatively ，and content of schisantherin A in S. sphenanthera is high relatively. Schizandrol A ，schizandrol B and schisandrin B were not detected in S. sphenanthera . The 3-color spatial value of S. sphenanthera and S. chinensis are different ，that is ，the brightness of S. chinensis is small and the color is slant black ，while the color of S. sphenanthera is slant red and yellow. The contents of active components of S. sphenanthera and S. chinensis is related to the surface 3-color spatial values ，that is ，the darker the color is ，the weaker the red degree is ，and the higher the contents of schizandrol A ，schizandrol B ，schisandrin B and schisandrin C are ；the brighter the surface color is ，the stronger the red degree is ，and the higher the contents of schisandrin A and schisantherin A are. The established content determination method is precise and stable ，and can be used for the content determination of S. sphenanthera and S. chinensis . The color qualitative identification model can be used for the identification of S. sphenanthera and S. chinensis .

OBJECTIVE： To prepare Resveratrol-hydroxypropyl-β-cyclodextrin-chitosan sustained-release pellets （RES-HP-β- CD-Chitosan）， and to characterize it. METHODS： Resveratrol raw material， HP-β-cyclodextrin and chitosan were collected with ratio of 1 ∶ 7 ∶ 0.25. Resveratrol-HP-β-cyclodextrin inclusion compound were prepared by solvent method， and then added into chitosan， RES-HP-β-CD-Chitosan were prepared by spray drying method. Particle size of prepared sustained-released pellets were observed by optical microscope. X-ray， DSC， IR and SEM were used to characterize RES-HP-β-CD-Chitosan. The contents of resveratrol in prepared sustained-released pellets were determined by UV spectrum， and drug-loading amount and encapsulation efficiency were calculated. RESULTS： Particle size of prepared RES-HP-β-CD-Chitosan was （2.23±0.35） μm （n＝300）. Characterization results show that RES-HP-β-CD-Chitosan was spherical in shape； shrinkage was found on the surface of microspheres， and resveratrol was included in HP-β-cyclodextrin in molecule or amorphous state. Drug-loading amount of prepared RES-HP-β-CD-Chitosan was 11.67％ （n＝3）， encapsulation efficiency was 96.27％ （n＝3）. CONCLUSIONS： RES-HP-β-CD- Chitosan is prepared successfully.

OBJECTIVE： To investigate in vitro release rate and in vivo pharmacokinetics of Resveratrol/hydroxypropyl-β- cyclodextrin/chitosan sustained-release pellets （RES/HP-β-CD/Chitosan） in rats. METHODS： In vitro release rate of RES raw materials， RES-HP-β-CD complexes （RES/HP-β-CD） and RES/HP-β-CD/Chitosan in water within 12 h were investigated by paddle method. The pharmacokinetic characteristics of RES raw materials， RES/HP-β-CD and RES/HP-β-CD/Chitosan were compared within 720 min after intragastric administration. RESULTS： Compared with RES raw materials， in vitro release rate of RES/HP-β-CD was increased significantly， and 120 min accumulative release rate reached 87％. Compared with RES/HP-β-CD， in vitro release rate of RES/HP-β-CD/Chitosan were relieved significantly； release time prolonged significantly； 12 h accumulative release rate was 72％. The pharmacokinetic parameters of RES raw materials， RES/HP-β-CD and RES/HP-β-CD/Chitosan included that cmax were 473.3， 2 492.2， 590.5 ng/mL； t1/2 were 2.6， 0.5， 4.6 h； AUC0-12 h were 514.7， 824.6， 2 778.5 ng·h/mL. Compared with RES raw materials， relative bioavailability of RES/HP-β-CD and RES/HP-β-CD/Chitosan were 172.5％ and 540.0％. CONCLUSIONS： RES/HP-β-CD/Chitosan shows good sustained-release effect， and its bioavailability is significantly higher than that of RES raw materials， RES/HP-β-CD.

Objective:To observe the effects of Deoxyschisandrin on hemorrheology and coagulation function in ulcerative colitis (UC)mice.Methods:Trinitrobenzenesulfonic acid (TNBS)induced UC mice model was prepared.And then UC mice were randomly divided into model group,positive control group,high,medium,and low dose groups of deoxyschisandrin (80,40,20mg/kg),and in addition,a normal control group was set up.There were 10 mice in each group respectively.UC mice were intragastricly administrated with different concentration of deoxyschisandrin in medication group,or with equal volume distilled water in normal group or model group,respectively.The blood viscosity was determined by blood rheometer,and the bleeding time (BT)and the clotting time (CT)were also observed through the methods of tail cutting and blood coagulation in glass plates accordingly.Results:Compared with model group,the BT (P ＜ 0.01)and CT (P ＜ 0.05)were significantly prolonged,and the blood viscosity was decreased obviously (P ＜ 0.05) in UC mice after administrated with different concentration deoxyschisandfin for 14 days.And the effects in high dose group were strongest and similar to those in the positive group.Conclusions:Deoxyschisandrin can improve hemorrheology and coagulation function in UC mice.

Ardipusilloside-I is a natural triterpenoid saponin, which was isolated from Ardisia pusilla A. DC. The aim of the study was to evaluate the stimulation of ardipusilloside-I on gastrointestinal motility in vitro and in vivo. The experiment of smooth muscle contraction directly monitored the contractions of the isolated jejunal segment (IJS) in different contractile states, and the effects of ardipusilloside-I on myosin were measured in the presence of Ca²⁺-calmodulin using the activities of 20 kDa myosin light chain (MLC₂₀) phosphorylation and myosin Mg²⁺-ATPase. The effects of ardipusilloside-I on gastro emptying and intestinal transit in constipation-predominant rats were observed, and the MLCK expression in jejuna of constipated rats was determined by western blot. The results showed that, ardipusilloside-I increased the contractility of IJS in a dose-dependent manner and reversed the low contractile state (LCS) of IJS induced by low Ca²⁺, adrenaline, and atropine respectively. There were synergistic effects on contractivity of IJS between ardipusilloside-I and ACh, high Ca²⁺, and histamine, respectively. Ardipusilloside-I could stimulate the phosphorylation of MLC₂₀ and Mg²⁺-ATPase activities of Ca²⁺- dependent phosphorylated myosin. Ardipusilloside-I also stimulated the gastric emptying and intestinal transit in normal and constipated rats in vivo, respectively, and increased the MLCK expression in the jejuna of constipation-predominant rats. Briefly, the findings demonstrated that ardipusilloside-I could effectively excite gastrointestinal motility in vitro and in vivo.
Animals ; Ardisia ; Atropine ; Blotting, Western ; Epinephrine ; Gastric Emptying ; Gastrointestinal Motility* ; Histamine ; In Vitro Techniques ; Muscle, Smooth* ; Myosin Light Chains* ; Myosin-Light-Chain Kinase* ; Myosins* ; Phosphorylation* ; Rats ; Saponins

OBJECTIVETo study the correlation between ITS sequence of Arctium lappa and Fructus Arctii quality of different origin.

METHODThe samples of Fructu arctii materials were collected from 26 different producing areas. Their ITS sequence were determined after polymerase chain reaction (PCR) and quality were evaluated through the determination of arctiin content by HPLC. Genetic diversity, genotype and correlation were analyzed by ClustalX (1.81), Mage 4.0, SPSS 13.0 statistical software.

RESULTITS sequence of A. was obtained from 26 samples, and was registered in the GenBank. Corresponding arctiin content of Fructus arctii and 1000-grain weight were determined. A. lappa genotype correlated with Fructus arctii quality by statistical analysis.

CONCLUSIONThe research provided a foundation for revealing the molecular mechanism of Fructus arctii geoherbs.

Arctium ; chemistry ; genetics ; Computational Biology ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; standards ; Furans ; analysis ; Genotype ; Glucosides ; analysis ; Polymorphism, Genetic ; genetics ; Quality Control ; Sequence Analysis, DNA ; Software

OBJECTIVETo identify the traditional medicine Fructus Arctii from its adulterants by ITS.

METHODTwenty-six samples of the different Fructus Arctii materials and 10 samples of the adulterants of the fruits of A. tomentosum, Onopordum acantium, Silybum marianum, and Amorpha fruticosa were collected. The total DNA of the samples were extracted, amplified cloned and sequenced.

RESULTITS sequences were obtained from 26 samples respectively, there were Fructus Arctii 641 bp, A. tomentosum 641 bp, Onopordum acantium 639 bp, Silybum marianum 630-631 bp, Amorpha fruticosa 625-626 bp, which were registered in the GenBank. The similarity in ITS sequences between Fructus Arctii and the adulterants was less than 95%. In contrast, the similarity between any pair of Fructus Arctii was greater than 99%. The similarity was 98.29%-99.22% between Fructus Arctii and A. tomentosum. Phylogeny tree reconstruction using UPGMA analysis based on ITS nucleotide sequences can effectively distinguish Fructus Arctii from adulterants.

CONCLUSIONITS sequences can be used as a reliable molecular marker for the identification of Fructus Arctii.

Arctium ; genetics ; Cell Nucleus ; genetics ; DNA, Ribosomal Spacer ; Drug Contamination ; prevention & control ; Genetic Markers ; Phylogeny ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

The roots, barks, branches and pericarps of Juglans mandshurica were used as folk medicine in China and reputed for its treatment of several cancers, such as gastric cancer, liver cancer and leukemia. The extracts of the roots, branches, leaves and pericarps of J. mandshurica have been experimentally proved to show anti-tumor activities. Tannins, which exhibited antioxidant and anti-tumor activities, were the main constituents in J. mandshurica. In this paper, a simple spectrophotometric method was developed for the determination of total tannins in the roots, branches, leaves and pericarps of J. mandshurica collected in Dalian and Anshan of Liaoning Province. Gallic acid was used as standard compound and the content of total tannins was calculated as gallic acid equivalent. As a result of the method validation, a good linearity (r = 0.9997, n = 5) and a high recovery of gallic acid (99.02%, RSD 3.7%, n = 9) was achieved. Eight samples including four parts of J. mandshurica collected in two places were analyzed for their total tannins with the established method. In the corresponding parts of J. mandshurica, except the pericarps, the contents of total tannins showed no significant difference between samples collected in Dalian and Anshan, while the content of total tannins in different parts of J. mandshurica were significantly different. The average content of total tannins in the roots, branches, leaves and pericarps of samples collected in Dalian and Anshan was 45.66, 23.40, 58.24, 3.58 mg g(-1), respectively.
Juglans ; chemistry ; Plant Extracts ; analysis ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Tannins ; analysis