Objective To establish the ultra-high performance liquid chromatography(UPLC)chromatographic fingerprint of Notopterygii Rhizoma et Radix;To determine the contents of ferulic acid,nodakenin,ammijin,notopterol,isoimperatorin and volatile oil of Notopterygii Rhizoma et Radix from different producing areas;To provide reference for quality evaluation of Notopterygii Rhizoma et Radix.Methods Waters BEH C18 chromatographic column(2.1 mm×150 mm,1.7 μm)was used,with mobile phase acetonitrile-0.02%formic acid aqueous solution gradient elution,flow rate 0.25 mL/min,column temperature 25℃,detection wavelength 330 nm,injection volume 2 μL.UPLC fingerprints of 25 batches of Notopterygii Rhizoma et Radix were established,and the similarity analysis and chemometrics analysis were carried out.The contents of ferulic acid,nodakenin,ammijin,notopterol and isoimperatorin were determined simultaneously,and the contents of volatile oil was determined by steam distillation method.Results Totally 23 common fingerprint peaks were calibrated,11 known components were identified.According to the results of the cluster analysis and principal component analysis,25 batches of Notopterygii Rhizoma et Radix samples were divided into 3 categories,and the 6 potential differential components were screened out by orthogonal partial least squares-discriminant analysis(OPLS-DA).The results showed that the contents of notopterol and volatile oil from Sichuan Province were higher than those from Gansu Province and Qinghai Province.Conclusion The method established in the study is accurate and reliable,which can provide scientific basis and reference for the quality evaluation and control of Notopterygii Rhizoma et Radix.

Objective To investigate the effect and mechanism of Shuangzhu Kangxian Prescription(Astragali Radix,bran-fried Atractylodis Macrocephalae Rhizoma,vinegar-prepared Rhizoma Curcumae,Bupleuri Radix,Salviae Miltiorrhizae Radix et Rhizoma,Litchi Semen)on improving hepatitic fibrosis induced by carbon tetrachloride(CCl4)in rats based on the Wnt/β-catenin signaling pathway.Methods The rat model of hepatitic fibrosis was replicated by subcutaneous injection of 3.0 mL·kg-1 40%CCl4 twice a week for 8 weeks.SD rats were randomly divided into blank control group(n=8),model group(n=7),positive control group(n=7,intragastric administration of 43.19 mg·kg-1 silymarin),low-dose Chinese medicine group(n=6,intragastric administration of 4.3 g·kg-1Shuangzhu Kangxian Prescription),medium-dose Chinese medicine group(n=6,intragastric administration of 8.6 g·kg-1Shuangzhu Kangxian Prescription),high-dose Chinese medicine group(n=7,intragastric administration of 17.2 g·kg-1Shuangzhu Kangxian Prescription).The continuous intragastric administration was given once a day for 4 consective weeks,in addition to the blank control group,the other groups continued to be subcutaneously injected with CCl4 at the same time.The pathological changes of liver tissue were observed by HE and Masson staining.The levels of serum type Ⅳ collagen(Col Ⅳ),type Ⅲ procollagen(PC Ⅲ),hyaluronic acid(HA)and laminin(LN)were detected by ELISA.The protein expression levels of Wnt1,β-catenin and PPAR-γ in liver tissue were detected by Western Blot.The mRNA expression levels of Wnt1,β-catenin and PPAR-γ in liver tissue were detected by qPCR.Results Compared with the blank control group,the liver of the model group showed partial necrosis of liver cells,the normal hepatic lobule structure was destroyed,the arrangement of liver cell cords was disordered,the central vein or portal area was enlarged,and a large number of inflammatory cells infiltrated to form bridging necrosis.The collagen fibers in the central vein or portal area of the liver proliferated significantly,forming fibrous septa,and the fibrosis score were significantly increased(P＜0.05).The levels of serum Col IV,PC Ⅲ,HA and LN were significantly increased(P＜0.05).The protein and mRNA expression levels of Wnt1 and β-catenin in liver tissue were significantly increased(P＜0.05),and the protein and mRNA expression levels of PPAR-γ were significantly decreased(P＜0.05).Compared with the model group,the steatosis of hepatocytes in the positive control group and the medium-and high-dose groups of Chinese medicine was improved,and the necrosis and inflammatory cell infiltration were reduced.The degree of hepatitic fibrosis was improved,the liver collagen fibers were reduced,and the fibrosis score was significantly reduced(P＜0.05).The levels of serum Col Ⅳ,PC Ⅲ,HA and LN were significantly decreased(P＜0.05).The protein and mRNA expression levels of Wnt1 and β-catenin in liver tissue were significantly decreased(P＜0.05),and the protein and mRNA expression levels of PPAR-γ were significantly increased(P＜0.05).Conclusion Shuangzhu Kangxian Prescription has the effect of anti CCl4-induced hepatitic fibrosis in rats,and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway and up-regulation of PPAR-γ expression.

Objective:To establish ultra performance liquid chromatography (UPLC) characteristic chromatogram of Pulsatilla chinensis; To provide reference for the origin identification and quality control of Pulsatilla chinensis. Methods:UPLC Method was adopted. The determination was performed on a column of Agilent SB C18 (2.1 mm×100 mm, 1.8 μm) . The mobile phase was acetonitrile-methanol (2:1) -0.1% phosphoric acid solution by fradient elution at a flow rate of 0.30ml/min. The column temperature was 30 ℃. The detection wavelength was 215 nm. The injection volume was 2 μl. The common counterfeit products and medicinal herbs of Pulsatilla chinensis from different areas were evaluated by comparison of characteristic chromatogram, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Results:There were 9 characteristic peaks in the characteristic chromatogram of Pulsatilla chinensis, and 8 common peaks were identified by high resolution mass spectrometry and comparison of reference materials. Through PCA analysis, it was possible to clearly distinguish the medicinal herbs of Pulsatilla chinensis from different areas. Combined with OPLS-DA analysis, it was found that peak 2, peak 3, peak 6 were the main markers of Pulsatilla chinensis from different producing areas. Conclusion:The established method has good specificity, repeatability and durability, and it can effectively distinguish the common counterfeits of Pulsatilla chinensis, and provide the basis of quality control and selection of origin for Pulsatilla chinensis.

Gastric cancer is a common malignant tumor of the gastrointestinal tract， with the pathogenesis remains to be fully elucidated. Although surgery， radiotherapy， chemotherapy， targeted therapy， and immunotherapy have demonstrated obvious clinical efficacy in the treatment of gastric cancer， the patients suffer from complications and adverse effects. Basic experiments and clinical studies have proved that Chinese medicine can treat gastric cancer in a multi-component and multi-target manner， the mechanisms of which remain to be deciphered. Therefore， the mechanism of Chinese medicine against gastric cancer needs to be unveiled by network pharmacology and tools of molecular biology. According to Chinese medicine， the occurrence of gastric cancer is mainly attributed to liver Qi stagnation， phlegm stasis and Qi stagnation， body fluid deficiency and heat accumulation， deficiency of healthy Qi， and cancer toxin accumulation. According to the available literature， herbal compound formulas such as Sancao Tiaowei decoction， Xiaojianzhong decoction， and Yiqi Huayu Jiedu decoction focus on tonifying， clearing heat， and detoxifying， while herbal active components are mainly insecticidal， heat-clearing， blood-activating， stasis-removing， and Qi-regulating drugs. The therapeutic effects of these Chinese medicines are consistent with the etiology and pathogenesis of gastric cancer. It has been demonstrated that Chinese medicines play a role in promoting apoptosis and autophagy， blocking cell cycle， and reversing cellular drug resistance to treat gastric cancer by regulating phosphatidylinositol-3 kinase/protein kinase B （PI3K/Akt）， mitogen-activated protein kinase （MAPK）， nuclear factor-kappa B （NF-κB）， transforming growth factor-β （TGF-β）/Smad， Wnt/β-catenin， and Hedgehog signaling pathways， while there is a lack of systematic understanding. By systematically summarizing the signaling pathways related to the regulation of gastric cancer by Chinese medicine， this study aims to clarify the molecular mechanisms of Chinese medicine against the development， invasion， and metastasis of gastric cancer， with a view to providing new targets， perspectives， and ideas for the treatment of gastric cancer and promoting the modernization of traditional Chinese medicine.

ObjectiveTo study the effect of total flavone of Litchi Semen （TFL） on proliferation， apoptosis， migration， and invasion of hepatoma cells HepG2. MethodMethyl thiazolyl tetrazolium colorimetric （MTT） assay was used to detect the effect of different-dose TFL and cisplatin on the proliferation of HepG2 cells. TdT-mediated dUTP nick-end labeling （TUNEL） assay was used to detect the effects of low， medium， and high-dose （70， 140， 210 mg·L^-1） of TFL and cisplatin （60 mg·L^-1） on the apoptosis of HepG2 cells， thus selecting the optimal dose of TFL for the follow-up experiment. HepG2 cells were divided into a blank group， a TFL group （140 mg·L^-1）， a TFL+XL019 group （140 mg·L^-1 TFL+0.5 μmol·L^-1 XL019）， and a TFL+TPI-1 group （140 mg·L^-1 TFL+1 μmol·L^-1 TPI-1）. The effect of TFL on migration and invasion of HepG2 cells were examined by wound healing test and Transwell invasion assay， and the effect of TFL on the expression of epithelial-mesenchymal transition （EMT） marker in HepG2 cells were examined by cell immunofluorescence assay. Western blot was used to detect the expression of key proteins in Janus kinase 2（JAK2）/signal transducer and activator of transcription 3 （STAT3） signaling pathway after the intervention by TFL. ResultMTT assay showed that the proliferation of HepG2 cells was significantly inhibited by TFL and cisplatin at 24 and 48 h as compared with blank group （P<0.01）， and the half maximal inhibitory concentration （IC₅₀） of TFL on HepG2 cells was （136.7±2.40） mg·L^-1 at 24 h and （106.8±1.11） mg·L^-1 at 48 h. The IC₅₀ of cisplatin on HepG2 cells was （58.48±2.04） mg·L^-1 at 24 h and （5.15±0.56） mg·L^-1 at 48 h. The results of TUNEL assay showed that TFL induced apoptosis of HepG2 cells. The optimal dose of TFL was 140 mg·L^-1. The results of wound healing test showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group significantly inhibited the migration of HepG2 cells （P<0.05， P<0.01）. As compared with the TFL group， the inhibitory effect of the TFL+XL019 Group was significantly increased （P<0.05）， while that of the TFL+TPI-1 group was significantly decreased （P<0.01）. The Transwell invasion assay showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group significantly inhibited the invasion of HepG2 cells （P<0.01）. As compared with the TFL group， the inhibitory effect of the TFL+XL019 group was significantly increased （P<0.05）， while that of the TFL+TPI-1 group was significantly decreased （P<0.01）. The results of immunofluorescence showed the intervention of TFL up-regulated the expression of E-cadherin， and down-regulated the expression of Vimentin in HepG2 cells， which was stronger in the TFL+XL019 group and weaker in the TFL+TPI-1 group. The results of Western blot showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group did not affect the expression of JAK2 or STAT3 protein， but significantly decreased the expression levels of phosphorylatied （p）-JAK2 and p-STAT3 （P<0.05， P<0.01）. As compared with the TFL group， the expression levels of p-JAK2 and p-STAT3 in the TFL+XL019 group were significantly decreased （P<0.01）， while those in the TFL+TPI-1 group were significantly increased （P<0.01）. Compared with the blank group， the TFL group significantly increased the expression level of Src-homology domain 2 containing protein tyrosine phosphatase-1（SHP-1） with sh2 domain （P<0.01）. ConclusionTFL has the effects of inhibiting the proliferation， promoting apoptosis of HepG2 cells， and reversing the EMT process of HepG2 cells to reduce the migration and invasion， which are presumably related to the activation of SHP-1 by TFL to block JAK/STAT3 signaling pathway.

OBJECTIVE To establi sh the fingerprint of Cnidium monnieri and a method for the content determination of 4 kinds of coumarins. METHODS Ultra-high performance liquid chromatography （UPLC） method was adopted to establish the fingerprints of 21 batches of C. monnieri ； their similarities were evaluated with Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）；common peaks were identification by comparison with reference substance. Using 10 common peak areas as variables ，cluster analysis was performed for 21 batches of C. monnieri by the method of between groups. The relative correction factors of xanthotoxin ，bergapten and imperatorin were calculated by the same UPLC method with osthole as the internal reference. The contents of them were calculated by quantitative analysis of multi-components by single marker （QAMS），and compared with the results of external standard method. RESULTS Totally 10 common peaks were identified in the fingerprints of 21 batches of C. monnieri ；the similarities ranged from 0.997 to 1.000. Peak 4 was identified as xanthotoxin ，peak 8 as bergapten ，peak 9 as imperatorin and peak 10 as osthole. A total of 21 batches of samples were divided into 3 categories，of which S 7 was clustered into one category ，S14 was clustered into one category ，and the other 19 batches were clustered into one category. The relative deviations of the contents of xanthotoxin ，bergapten and imperatorin determined by QAMS and external standard method were in the range of 0.88％ -1.07％ ，2.22％ -2.29％ ，0.67％ -2.93％ ，respectively. CONCLUSIONS UPLC fingerprint of C. monnieri is successfully established ，and QAMS method for content determination of 4 coumarins is also established.

OBJECTIVE：To establish the UPLC fingerprint of Polygonum cuspidatum ，and to determine the contents of four active ingredients and to provide reference for the quality evaluation of P. cuspidatum . METHODS ：The determination was performed on Waters BEH C 18 column（100 mm×2.1 mm，1.7 μm）with mobile phase consisted of acetonitrile- 0.2％ formic acid （gradient elution ）at flow rate of 0.4 mL/min. The column temperature was 40 ℃，and detection wavelength was 290 nm. The sample size was 1 μL. The fingerprints were evaluated by similarity calculation，cluster analysis and orthogonal partial least square discriminant analysis （OPLS-DA）. Using polydatin as internal standard ，relative calibration factors of resveratrol ，emodin-8-O- β-D-glucoside and emodin were determined to develop a method of QAMS. The contents of 4 above components in 15 batches of P. cuspidatum were calculated by relative calibration factors. The results of QAMS were compared with those of external standard. RESULTS：UPLC fingerprints of 15 batches of P. cuspidatum were established ，and 12 common peaks were confirmed. Five components were identified ，i.e. polydatin ，resveratrol，emodin-8-O-β-D-glucoside，emodin，emodin methyl ether. The fingerprint similarity of 15 batches of P. cuspidatum was in the range of 0.865-0.976. According to cluster analysis ，15 batches of P. cuspidatum were classified into 4 categories，showing certain regularity of origin. Seven markers were identified by OPLS-DA method. The order of difference significance was peak 7＞emodin-8-O-β-D-glucoside＞resveratrol＞peak 8＞polydatin＞peak 1＞ peak 10. The relative deviation among the contents of resveratrol ，emodin-8-O-β-D-glucoside and emodin in 15 batches of P. cuspidatum determined by QAMS and external standard method was less than 5.0％，indicating that there was no significant difference between the two methods. CONCLUSIONS ：UPLC fingerprint combined with QAMS method is convenient and reliable for the quality evaluation of P. cuspidatum ；the quality of P. cuspidatum produced in Chongqing and Anhui province is better.

OBJECTIVE：To establish UPLC fingerpri nt of Ligusticum sinense ，Ligusticum jeholense and Conioselinum vaginatium，and to conduct their chemometrics analysis so as to provide reference for the identification of Rhizoma Ligustici from different origins. METHODS ：UPLC method combined with Similarity Evaluation System of TCM Chromatographic Fingerprints （2012 edition） were used to establish the fingerprints of Rhizoma Ligustici from different origins. Chromatographic peak identification and similarity evaluation were carried out. Cluster analysis （CA），principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA）were performed to analyze Rhizoma Ligustici from different origins，and screen differential components. RESULTS ：Totally 13，11，11 characteristic peaks were identified in UPLC fingerprints of L. sinense ，L. jeholense and C. vaginatium ，respectively. Similarity evaluation showed that the similarity between C. vaginatium and L. jeholense were 0.312-0.541；that between C. vaginatium and L. sinense were 0.324-0.682；that between L. sinense and L. jeholense were 0.312-0.671，indicating there was great difference among Rhizoma Ligustici from different origins. CA，PCA and OPLS-DA showed that Rhizoma Ligustici from different origins were clustered into each category respectively ； chemical components represented by peak 10，peak 13，peak 12，peak 7 and peak 6 were differential components for Rhizoma Ligustici from 3 origins. CONCLUSIONS ：The study establishes UPLC fingerprint of Rhizoma Ligustici from different origins ， and screens 5 differential components ，which can be used to identify Rhizoma Ligustici from different origins.

Objective: To explore the clinical effect of a guided resin cementation technique on vertical food impaction symptoms and to provide a new method for the treatment of vertical food impaction. Methods : Treatment of 76 patients with vertical food impaction with guided resin cementation was performed. A specially fabricated contact shaping wire was used to aid the shaping of the contact. Cementation was applied under a rubber dam with the total-etch technique with flowable composite resin. Patient subjective perception was recorded after treatment (i.e., “totally relieved”=3, “significantly improved”=2, “slightly improved”=1 and “no change”=0). Follow-up visits lasted for one year. Scores of 1 to 3 were recorded as effective. The efficiency rates at different times were calculated. Results: Patient subjective perceptions scored 2.47, 2.21, 1.79, 1.30 and 0.97 on the day immediately after and 1, 3, 6, and 12 months after treatment, respectively. There were significant differences among scores at each time point (P<0.01). The Efficacy rate reached 91.78% immediately after treatment and was sustained above 50% within half a year. Management of resin debonding or fracture successfully relieved the symptoms again. Conclusion The guided resin cementation technique relieves vertical food impaction symptoms immediately, and the effect of the guided resin cementation technique is maintained for a short period of time. Management of resin debonding or fracture helps consolidate treatment outcomes.

Aim To investigate the effects of ursolic acid from loquat leaves on proliferation inhibition and expression of PPAR-γ 、TGF-β1 in rat hepatic stellate cells, so as to explore the mechanism of anti-hepatic fibrosis of UA.Methods HSC-T6 cells were randomly divided into blank group, rosiglitazone control group, the low, medium and high concentration of UA group to detect the cell proliferation inhibition by CCK-8 after 24,48,72 h.The content of type collagen Ⅰ in cell culture supernatant of each group was examined by ELISA.The expression of PPAR-γ mRNA, TGF-β1 mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR.Effects on HSC-T6 PPAR-γ and TGF-β1 protein from each group were detected by immunocytochemical method.Results CCK-8 results showed that the inhibitory rate of UA on cell proliferation increased with the prolongation of drug action time(P<0.01).ELISA results showed that with the increase of the concentration of UA, the content of type Ⅰ collagen content decreased(P<0.01).Real-time PCR results showed that with the increase of the concentration of UA, and the expression of PPAR-γ mRNA increased, the expression of TGF-β1 mRNA decreased(P<0.01).The results of immunocytochemistry showed that the expression of PPAR-γ protein was increased, and the expression of TGF-β1 protein was decreased with the increase of the concentration of UA(P<0.01).All effects mentioned above were dose-dependent.Moreover, the effects in the high concentration groups were stronger than those in control group.Conclusion UA can inhibit the proliferation of HSC-T6 cells, Which may be associated with the up-regulation of PPAR-γ expression and the down-regulation of TGF-β1 expression.