The dynamic metabolic profile of Qi-Yu-San-Long decoction in rat urine using UPLC-QTOF-MSE coupled with a post-targeted screening strategy / 药物分析学报

Qi-Yu-San-Long decoction(QYSLD)is a traditional Chinese medicine that has been clinically used in the treatment of non-small-cell lung cancer(NSCLC)for more than 20 years.However,to date,metabolic-related studies on QYSLD have not been performed.In this study,a post-targeted screening strategy based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight full infor-mation tandem mass spectrometry(UPLC-QTOF-MSE)was developed to identify QYSLD-related xeno-biotics in rat urine.The chemical compound database of QYSLD constituents was established from previous research,and metabolites related to these compounds were predicted in combination with their possible metabolic pathways.The metabolites were identified by extracted ion chromatograms using predicted m/z values as well as retention time,excimer ions,and fragmentation behavior.Overall,85 QYSLD-related xenobiotics(20 prototype compounds and 65 metabolites)were characterized from rat urine.The main metabolic reactions and elimination features of QYSLD included oxidation,reduction,decarboxylation,hydrolysis,demethylation,glucuronidation,sulfation,methylation,deglycosylation,acetylation,and associated combination reactions.Of the identified molecules,14 prototype compounds and 58 metabolites were slowly eliminated,thus accumulating in vivo over an extended period,while five prototypes and two metabolites were present in vivo for a short duration.Furthermore,one pro-totype and five metabolites underwent the process of"appearing-disappearing-reappearing"in vivo.Overall,the metabolic profile and characteristics of QYSLD in rat urine were determined,which is useful in elucidating the active components of the decoction in vivo,thus providing the basis for studying its mechanism of action.

Immune dysfunction of regulatory B cells and lymphocyte subsets in children with primary immune thrombocytopenia / 中国医师杂志

Objective:To investigate the changes of regulatory B cells (Bregs) and lymphocyte subsets in children with primary immune thrombocytopenia (ITP), and to analyze their relationship with ITP duration and remission time.Methods:Proportion of different immune cell subsets were detected in the peripheral blood of 88 ITP children before treatmen, 84 ITP children after treatmen and 45 normal controls by flow cytometry. The treatments included glucocorticoids, intravenous IgG as first-line treatment. The changes of lymphocyte subsets in ITP children with different remission time after treatment were compared.Results:The Th, natural killer (NK), Breg cell counts and Th/Tc ratio in all children with ITP were significantly lower than those in the control group ( P<0.05), while the Tc and CD19 + B cells were significantly higher than those in the control group ( P<0.05). The Breg cell count in the persistent and chronic ITP groups was significantly lower than that in the newly diagnosed ITP ( P<0.05). A total of 84 children with ITP were relieved after treatment: 58 cases (69.1%) achieved long-term remission, and 26 cases (30.9%) had short-term remission. The Breg and lymphocyte subsets in the short-term remission ITP group after treatment were not significantly different from those before treatment, but there were still significant differences from the control group ( P<0.05); The Breg and lymphocyte subpopulations in the long-term remission ITP group after treatment were not significantly different from those in the control group, but compared with those before treatment, other cell subpopulations except Tc were significantly increased ( P<0.05); The NK cells and Breg cells in the long-term remission ITP group were significantly higher than those in the short-term remission ITP group ( P<0.05). Conclusions:The Bregs in peripheral blood of long-duration ITP children significantly decrease. After treatment, the Bregs in ITP children with long-term remission increase significantly, indicating that the restoration of the immune mechanism of Bregs may be related to the long-term remission of children with ITP.

Anti-leukemia effect of icaritin and its mechanisms on cell line of children relapsed acute lymphoblastic leukemia / 中国医师杂志

Objective:To investigate the anti-leukemia effect of icaritin on 697 cells of children relapsed acute lymphoblastic leukemia in vitro, and to analyze its possible mechanism. Methods:697 cells were cultured in vitro and divided into control group and icariin groups (5, 10, 20 μmol/L). After treated with icaritin, the cell proliferation was detected with cell counting kit-8 (CCK-8) method, and apoptosis was measured with Hoechst 33258 staining and Flow Cytometry Assay. Expression of Bcl-2, Cyt-C, Bax and Caspase3 proteins in 697 cell were detected by Western blot. Results:After treatment with various concentrations (5-20 μmol/L) of icaritin for different time-points (24, 48, 72 h), the growth inhibition rate exhibited a dose and time-dependent manner ( r=0.76, r=0.89); the apoptotic rate exhibited significant increase for 48 h ( P<0.05). Bcl-2 protein decreased significantly ( P<0.05), while Bax, Cyt-C and Caspase3 cleavage protein increased significantly in icaritin treated group for 48 h compared with control group ( P<0.05). Conclusions:Icaritin shows the effects of anti-leukemia by inducing apoptosis and inhibiting proliferation of 697 cell. Icaritin maybe induce apoptosis of 697 cell by down-regulation of Bcl-2 expression and up-regulation of the Bax, Cyt-C and cleaved Caspase 3 expression.

Changes of immunocyte subsets and their related factors in children with immune thrombocytopenia / 中国医师杂志

Objective To investigate the changes of T cells and dendritic cells (DCs) and their related factors in children with immune thrombocytopenia (ITP) before and after therapy,and to analyze their clinical significance.Methods T-cells and DCs subsets were determined by flow cytometry both in 64 children with ITP (ITP group) before and after therapy and the control group.The serum levels of interleukin (IL)-4,interferon-γ (IFN-γ),transforming growth factor beta 1 (TGF-β1),and IL-27 were detected by enzyme linked immunosorbent assay (ELISA).Results Treatments of glucocorticoid or IVIg were effective in 41 cases of 64 ITP children.Compared to the control group,helper T cells (Th),Th/suppressor T cells (Ts),T regulatory cells (Treg),plasmacytoid DC (pDC),pDC/myeloid DC (mDC),and TGF-β1 in ITP patient group before treatment were significantly lower,while IFN-γ and IFN-γ/IL-4 were significantly higher (P ＜0.05).In ITP group,Th,Th/Ts,Treg,pDC,pDC/mDC,TGF-β1,and IL-27 were significantly increased,while IFN-γ and IFN-γ/IL-4 were decreased in children with ITP after therapy and achieved response (P ＜ 0.05).However,there was no significant difference between before and after therapy in ITP children without treatment response (P ＞ 0.05).Conclusions T cells and DCs subsets disorder and abnormal cytokine levels are observed in children with ITP,which can be corrected by immunosuppressive therapy,indicating that Th1 overactivity and the decrease of Treg and pDC both in quantity and function may be related to the pathogenesis of ITP in children.

Determination of New Bistrifluron, Cyantraniliprole and SYP9080 Residues in Fruits and Vegetables by Liquid Chromatography-Tandem Mass Spectrometry / 分析化学

A liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of bistrifluron, cyantraniliprole and SYP9080 residues in fruits and vegetables. The analytes were extracted from samples with acetonitrile, and purified by dispersion solid phase extraction using C18 and primary secondary amine (PSA) as solid phase adsorbent, then analyzed by LC-MS/MS. Bistrifluron and SYP9080 were analyzed with negative ion multiple-reaction monitoring (MRM), cyantraniliprole was analyzed with positive ion MRM. The matrix-induced weakening effect was observed in the analysis of cyantraniliprole and SYP9080 in several samples including cucumber, apple and onion, and the weakening extent of the matrix-induced effect depended on the sample properties. But no matrix effect was found in the analysis of bistrifluron in several samples. The interference of matrix was reduced by using the matrix-matched calibration standards curve in the analysis of cyantraniliprole and SYP9080. Bistrifluron could be quantified by an external standard of the matrix-matched calibration standards or the pure solvent calibration standards. The linear range was from 0.2 μg/L to 100 μg/L for bistrifluron, cyantraniliprole and SYP9080 with the good correlation coefficients (r≥0.9990). The recoveries of cucumber, grape, apple and scallion added bistrifluron at 0.005-2 mg/kg were 79.8％-99.9％, with relative standard deviations (RSD) of 3.6％-9.8％. The limits of quantification (S/N=10) were 0.210 μg/kg, 0.160 μg/kg and 0.004 μg/kg, and limits of detection (S/N=3) were 0.064 μg/kg, 0.048 μg/kg and 0.001 μg/kg for bistrifluron, cyantraniliprole and SYP9080, respectively.

Research on relevance between mitochondrial ATP synthase and malignant tumor / 生物医学工程学杂志

Mitochondrial adenosine triphosphate (ATP) synthase is the key enzyme of mitochondrial oxidative phosphorylation reaction. The down-regulation of the mitochondrial ATP synthase is a hallmark of most human carcinomas, which is the embodiment of the bioenergetic signature of cancer in the performance of the decreased oxidative phosphorylation and increased aerobic glycolysis. Combining with the bioenergetic signature of cancer, studies showed that mitochondrial ATP synthase and multidrug resistance and adverse prognosis of tumor were closely related. Its mechanisms are related to post-transcriptional regulation of the ATP synthase, the hypermethylation of the ATP synthase gene and the inhibitor peptide of the mitochondrial ATP synthase, called ATP synthase inhibitory factor 1 (IF1). In this review, we stress the biological characteristics of mitochondrial ATP synthase and the relationship between ATP synthase and multidrug resistance and prognosis of Malignant tumor, in order to find a new way for tumor therapy.

Attitude to Career in Undergraduates of Rehabilitation Therapy / 中国康复理论与实践

Objective To investigate the attitude of career in undergraduates of rehabilitation therapy. Methods 41 undergraduate stu-dents were investigated with questionnaire. Results 14.6% of the students preferred rehabilitative therapeutist, 53.7% knew little about the profession, 31.7%selected this profession for employment, 22.0%loved rehabilitation work, 41.4%would take rehabilitation work for life. Conclusion It is important for school to help the students understanding the profession of rehabilitative therapeutist correctly.

Bone marrow mesenchymal stem cells cultured by umbilical cord blood serum versus adult autoserum in vitro / 中国组织工程研究

BACKGROUND:Studies concerning bone marrow mesenchymal stem cells (BMSCs) cultured by umbilical cord serum in vitro were hot topic to avoid the heterogenous serum rejection during BMSC culture and improve culture efficiency of BMSCs.OBJECTIVE:To compare biological feature of BMSCs by the umbilical cord serum and adult autoserum in vitro.DESIGN,TIME AND SETTING:The opening experiment was performed at the Laboratory of Cell Biology,Xiangya Second Hospital,Central South University,China from October 2006 to June 2008.MATERIALS:Sixteen bone marrow samples were provided by Xiangya Second Hospital,Central South University and Xiangtan Central Hospital of Hunan Province.METHODS:BMSCs were obtained from 16 fresh adult bone marrow by gradient centrifugation with Ficoll Paque,cultured with DMEM/F12 with 10% umbilical cord blood serum and 10% adult autoserum.The fourth passage was used in this study.The surface antigens of BMSCs were detected by flow cytometry.BMSCs could differentiate into ostenblasts and adipocytes under culture in conditioned medium for osteogeneais and adipogenesis and the differentiated BMSCs were identified by morphological observation,immunophenotype and immunochemical staining.MAIN OUTCOME MEASURES:The following parameters were measured:morphological observation;cell cycle and cell count;identification of surface antigen;observation of osteoblasts and adipocytea induced from BMSCs by immunochemical staining.RESULTS:The quantity and velocity of BMSCs in umbilical curd blood serum was better than those in adult autoserum (P ＜ 0.05)Only few cells were proliferating in both medium,BMSCs in S phase in umbilical cord blood serum was more than those in adult autoserum (P ＜ 0.05).The positive rate of CD29,CD73 and CD105 on BMSCs in umbilical cord serum (above 90%) was higher than those in adult autoserum (P ＜ 0.05),and the positive rates of CD31,CD34 and CD45 were lower than 2%,and the positive rate of CD31 was lower than those in adult autoserum (P ＜ 0.05).The positive rate of BMSCs differentiated into osteoblasts and adipocytes in umbilical cord blood serum was also higher than these in adult autoserum (P ＜ 0.05).CONCLUSION:Purity and differentiated potency of BMSCs in umbilical cord blood serum are better than those in the adult autoserum in vitro.The umbilical cord serum is more adapt to clinical application.