OBJECTIVE:To determine the content of chrysophanol in Diedazhitong cataplsam by HPLC.METHODS:The samples were separated on a Kromasasil C 18 ,methol-0.1%phosphoric acid(85∶15)was taken as the mobile phase with a flow rate at1.0ml/min and detection wavelength at254nm,the sample size was10?l.RESULTS:Good linear relationship was achieved when the concentration range of chrysophanol was0.6?g～4?g(r=0.9994);The average recovery of chrysophanol was99.23%(n=5,RSD=2.45%).CONCLUSION:This method is accurate,reliable,and reproducible.

Objectives To investigate the characteristics of changes in serum metabolic profile before and after resection of carcinoma tissues to establish a disease distinguishing model,to analyze the changing trend of characteristic metabolites,and to determine the molecular mechanism and potential clinical value of characteristic metabolic markers for HBV-related liver cancer.Methods Ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) was used to analyze the serum metabolites of 15 patients with hepatocellular carcinoma (HCC) before and after partial hepatectomy and on 25 healthy volunteers.The pattern recognition method and nonparametric test analyzes were used to analyze the data and to identify the specific metabolites and their changes after resection of carcinoma tissues.Results We established the principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) disease distinguishing model for HCC patients before and after operation as against the healthy volunteers.To distinguish between the liver cancer group and the normal control group,27 characteristic metabolites were selected from the patients before and after resection of carcinoma tissues.Eight moved towards the normal control after resection of carcinoma tissues.This indicated that liver carcinoma was an important impacting factor for these metabolites.Finally,7 metabolites were identified,and these metabolites had high diagnostic value as shown on ROC curves.Conclusions Through serum metabolic profiling of patients before and after resection of carcinoma tissues,a high correlation between metabolism and hepatocellular carcinoma was found.Researches on endogenous metabolites and pathways in liver diseases will provide a better understanding of the molecular mechanisms and provide further directions for clinical diagnosis and treatment.

[Objective] To investigate the variation of Q promoter (Qp) in nasopharyngeal carcinoma (NPC) cells, and to compare the existing two mutant sites [62 225 site(g→a)and 62 422 site (g→c) ] Qp in NPC cells with the Qp in B95.8 cell line in the functional and biological difference. [Methods] The Qp sequence was amplified in the samples from 29 cases of paraffin-embedded tissues of NPC suffers and 14 cases of peripheral blood of healthy adults by polymerase chain reaction (PCR) method (totally 43 cases). The point mutations on specified sites were analyzed and statistically compared from sequencing results. The sequences of variant and prototype Qp were amplified by PCR and cloned into luciferase reporter vector (pGL3-basic), then transfected into HaCat cells respectively. The transcriptional activity was compared between variant and prototype Qp using luciferase reporter system. The DNA binding affinity of mutant and prototype Qp to Sp1 was compared through chromatin immunoprecipitation (CHIP) method since mutation of nt 62 225 located in a Spl binding site. [Results] The mutation rate of Qp was significantly higher in NPC compared with healthy controls (P=0.039 5, <0.05), which suggested the variant Qp was closely associated with NPC. The transcription of the luciferase gene promoted by variant Qp was significant more than that of prototype Qp in transient transfection assay (2.5:1, P<0.05). The binding affinity of variant Qp to Sp1 was about 1.52 times higher than that of prototype Qp as determined by quantitative ChIP assay. [Conclusions] The transcriptional activity was enhanced in variant Qp in NPC cells compared with prototype, which possibly through the higher binding affinity to Sp1. We suggest that the mutated Qp may play an important role during the EBV infection and transformation of nasopharyngeal epithelium.

This study was aimed to establish a quantitative method for the determination of agarotetrol in agarwood to control its quality. The HPLC analysis was performed on a Diamonsil C18 column (4.6 mm í 250 mm, 5 μm), eluted with a mobile phase of water with 0.1% formic acid and acetonitrile in gradient mode with the flow rate of 0.7 mL·min-1. The detection wavelength was set at 252 nm. The column temperature was 30oC. The results showed that the separation of agarotetrol and adjacent peaks were more than 1.5, which achieved a baseline separation. A good lin-earity (r = 0.999 8) was observed in the range of 2.0-125.0 μg·mL-1, with the average recovery of 102.75%. The a-garotetrol cannot be less than 0.15% based on the data of Agarwood samples. It was concluded that the method was accurate and reliable to determine the content of agarotetrol, which can be used for the quality control of agarwood.

Objective:To investigate the functional status of dendritic cells(DCs)in oral leukoplakia(OLK)tissues.Methods:The expression of DC-specific markers CD1 a,CD209,CD1 23 and CD83 in 20 cases of OLK with abnormal dysplasia,1 0 with simple dys-plasia and 1 0 of normal oral mucosa tissues was examined by immunohistochemistry.Results:CD1 a and CD209 positive DCs were found in all cases.More CD1 a positive Langehans cells(LCs)in lamina propria were found in OLK with abnormal dysplasia than in normal o-ral mucosa and OLK with simple dysplasia(P <0.01 ).A great mount of CD209 positive stromal DCs were recruited in OLK.There was no CD83 positive and CD1 23 positive cell in normal oral mucosa,however,CD83 positive mature DCs and CD1 23 positive plasmacytoid DCs(PDCs)were observed in OLK(P <0.01 ).Conclusion:OLK is characterized by the recruitment of different subsets of DCs,the different DC subsets may play an important role in the development of OLK.

Objective To study the effects of Androsace umbellata extract on bone wound healing in rats.Methods A total of 32 rats were selected,and the rat femur bone trauma model was established.The Androsace umbellata was administrated to rats in treatment groups (including high-dose Androsace umbellata group and low-dose Androsace umbellata group,8 rats in each group)continuously for 10 days,while rats in the fake operation control group (8 rats)and bone trauma model group (8 rats) were treated with corresponding volume of solvent by body weight.The growth of body weight and wound healing of rats were recorded.The serum levels of calcium and phosphorus,activity of alkaline phosphatase (ALP),bone density and bone biomechanics were examined.The X-ray photograph was carried out to observe the effects of Androsace umbellata on bone wound healing,Results Compared with the bone trauma model group,serum levels of calcium and phosphorus,calcium-phosphorus product and activity of ALP were significantly increased in treatment groups,there were statistically significant differences (P＜0.05).Compared with the bone trauma model group,bone density of trauma place in the high-dose Androsace umbellata group was significantly decreased (P＜0.05),bending energy in the low-dose Androsace umbellata group was increased (P＜0.05),while no statistically significant difference was found in the other skeletal biomechanical properties (P＞0.05).The results of X-ray films indicated that the treatment groups shown better effects on bone wound healing compared with the bone trauma model group.Conclusion Androsace umbellata extract could effectively promote bone wound healing in rats.

OBJECTIVE To explore mecha-nisms of imperatorin on regulating P-glycoprotein(P-gp)in blood-brain barrier(BBB)based on net-work pharmacology combined with in vitro experi-ment.METHODS Drug targets were predicted using the Pharmapper and Swiss targets data-bases;disease targets were obtained through the Genecards database;intersections between drugs and disease targets were screened by Cytoscape software;the obtained core targets were used to construct protein-protein interaction(PPI)network,gene ontology(GO)functions,and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.The effects of imperatorin(20,50,100 μ mol·L-1)on P-gp activity were monitored in hCMEC/D3in vitro BBB model,and the effects of imperatorin on the expression of target proteins were verified using Western blot method.RESULTS 55 drug targets and 3102 disease targets were obtained from the network pharmacology screening,and 37 core targets were obtained after the combination.Enrichment analysis showed that core targets were closely related to chemical synaptic trans-mission regulation,neurotransmitter receptor activity,proteinkinaseregulationactivity,G protein-coupled receptor signaling pathway,neural active ligand receptor interaction pathway,PI3K-Akt sig-naling pathway,VEGF signaling pathway,etc..In vitro experimental validation suggested that all tested concentration groups of imperatorin signifi-cantly reduced the activity and expression of P-gp,which were achieved by significantly downregu-lating the phosphorylation levels of PI3K and Akt,and repressing the expression of VEGFR2 pro-tein.CONCLUSION Network pharmacology was used to predict the core targets and signaling pathways of imperatorin on regulating P-gp in BBB and relevant validation was conducted through in vitro experiments,providing a refer-ence basis for further exploration of the mecha-nisms of imperatorin on regulating P-gp in BBB.

Objective:To design an electronic nose that can detect and identify urinary volatile organic compounds (VOCs) as marker gases for bladder cancer.Methods:Isopropyl alcohol, ethylbenzene, acetic acid, and ammonia were selected as target gases, and 8 metal oxide gas sensors were used to construct sensor arrays for testing and collecting experimental data, and different characteristics were normalized. Recursive feature elimination (RFE) was used to select the best feature subset, and principal component analysis (PCA) and linear discriminant analysis (LDA) were further introduced to reduce the data dimension and facilitate visual analysis. In addition, three machine learning algorithms, including support vector machine (SVM), random forest (RF), and K-nearest neighbor (KNN), were combined to train and verify the model.Results:When the feature number was 12, the accuracy of the model classification had the best performance. The feature subset consisted of 5 differences, 5 sensitivities, and 2 integrals, and the data was reduced to 12 dimensions. Only PCA couldn’t distinguish the four gases. The LDA classification performance was significantly better than that of PCA, except that isopropyl alcohol and acetic acid had a small overlap area. LDA could distinguish ethylbenzene and ammonia from isopropyl alcohol and acetic acid; the sample points were gathered, which means the clustering performance was also better. The prediction accuracy of SVM, RF, and KNN was 0.85, 0.56, and 0.79, respectively. After model verification, the classification accuracy of PCA+SVM, LDA+RF, and LDA+KNN was 0.97, 0.94, and 0.97, respectively.Conclusions:An electronic nose was designed to detect and identify urinary VOCs marker gases for bladder cancer.

Lymphocytic hypophysitis(LYH) is a rare autoimmune inflammatory disorder of the pituitary gland, usually affecting young women in late pregnancy or postpartum period. To enhance the knowledge of LYH, herein we reported a case of LYH in a female during postpartum who presented with pituitary crisis.

Objective: To investigate the role of JAZF1 gene rearrangement in the diagnosis and differential diagnosis of endometrial stromal sarcomas by fluorescence in situ hybridization (FISH). Methods: JAZF1 gene rearrangement was analyzed by FISH in 129 cases of ESS diagnosed from January 2008 to December 2016 including 105 cases of low-grade endometrial stromal sarcoma (LG-ESS), 21 cases of high-grade endometrial stromal sarcoma (HG-ESS) and 3 cases of undifferentiated uterine sarcoma (UUS). Sixteen cases of the related tumours in uterus were also collected as control group. The results were compared with our previous studies of JAZF1/JJAZ1 fusion gene in ESS by RT-PCR. Results: Detection of JAZF1 gene rearrangement by FISH was successfully analyzed in 144 cases. JAZF1 gene alteration was detected in 63 cases, all of which were LG-ESS, with an overall positivity of 60.6% (63/104), while no JAZF1 gene rearrangement was found in all other cases. JAZF1 gene rearrangement was present in LG-ESS with classic histology (69.3%, 52/75), smooth muscle differentiation (2/10), sex cord-like differentiation (4/5), fibromyxoid change (1/5), clear cell change (0/1), skeletal muscle differentiation (0/1), and schwannoma-like palisading pattern (0/1). The different components in all the cases of LG-ESS with variant histology had the clonal origin, with or without JAZF1 gene alteration. Compared to the results of JAZF1/JJAZ1 fusion gene by RT-PCR, the positive rate of JAZF1 gene rearrangement in LG-ESS by FISH (61.9%, 26/42) was significantly higher than that of RT-PCR (30.0%, 12/40; P<0.01). Conclusions JAZF1 gene rearrangement is present only in LG-ESS, but not in HG-ESS, UUS or other related tumours in uterus. The frequency of JAZF1 gene rearrangement varies between classic LG-ESS and different morphologic variants. It is frequently, but not consistently, present in classic LG-ESS and less often positive in variant cases. The results of JAZF1 gene alterations in LG-ESS with different morphologic variants support the contention that the endometrial stromal and their variant morphologic components have the same clonal origin. Detection of JAZF1 gene rearrangement by FISH is very useful for the diagnosis and differential diagnosis of ESS.