The paper reports the establishment of a method for simultaneous determination of peimisine and sipeimine contents in Fritillaria walujewii Regel and Fritillaria pallidiflora Schrenk. The analyses were performed on an ultra-performance liquid chromatography with evaporative light scattering detection (UPLC-ELSD), equipped with a binary solvent manager, a sampler manager and a column compartment, and connected to Waters Empower 2 software. An Acquity UPLC BEH C18 column (100 mm x 2.1 mm, 1.7 microm) was used for all analysis. The investigated compounds were separated with gradient mobile phase consisting of acetonitrile-0.02% triethylamine-water. The temperature of sample manager was set at 25 degrees C. Drift tube temperature was 40 degrees C, and spray parameter was 40% with injection volume of 1 microL. The investigated compounds including peimisine and sipeimine had good linearity (r > or = 0.9991) over the tested ranges. The average recovery was 94.5% and 98.1% with RSD < or = 2.36%. The UPLC-ELSD method is simple, sensitive and accurate with good repeatability, which is available for quality control of F. walujewii Regel and F. pallidiflora Schrenk.

Objective To explore the effects of psoralen (PSO) and long wave ultraviolet A (PUVA) on apoptosis and expression of Fas in HL-60,K562 and NB4 leukemia cells.Methods The cells were incubated with PSO in different concentrations irradiated with or without UVA.The changes of ultrastructure of cells were observed under the electron microscope.The expression of Fas gene was detected by fluorescent quantitation PCR.The apoptosis ratio and the expression of Fas protein were detected through the flow cytometry.The factorial design and analysis of variance were used to analyze the interaction among the factors.Results There were obvious ultrastructure changes about apoptosis in leukemia cells after treated with PUVA.PSO,UVA and PUVA all increased the apoptosis ratio and expression of Fas gene and protein,and the effects of PUVA were stronger than the other two (P

0.05). Conclusion PASP is an actually non-poisonous material and not a mutagen, the result of the present paper may be taken as a scientific evidence for the safety of PASP to be used in industry water treatment.

To study the mechanism of therapeutie effect of compound "Huang Dai" tablet in acute promyelocytic leukemia, twenty one patients with acute promyelocytic leukemia were treated with compound "Huang Dai" tablet. Bone marrow cells of the patients were collected before treatment and after treatment, and membrane differentiating antigen (CD33 and CD11b), cell cycle and the apoptosis correlative protein(Fas and Apo 2.7 )were detected with flow cytometry. Results showed that the expression of CD33 was decreased and that of CD11b increased, G 2 /M phase of cell cycle was blocked, and the expressions of Fas and Apo 2.7 protein were increased significantly in all patents after treatment with compound "Huang Dai" tablet. These results suggested that the mechanism of therapeutic effect of compound "Huang Dai" tablet in acute promyelocytic leukemia was induction of both differentiation and apoptosis of leukemic cells, and cell apoptosis might be related to Fas and Apo2.7 protein.

OBJECTIVE: To explore the effects of psoralen (PSO) plus long-wave ultraviolet-A (PUVA) on apoptosis and expression of Fas ligand (FasL) in HL-60 leukemia cells. METHODS: The HL-60 cells were taken as the study objects and their apoptosis rates, ultrastructure changes and the expression of FasL were detected in order to observe the effects of PSO and ultraviolet-A (UVA) of wave length 360 nm. The factorial design and analysis of variance were used to analyze the interaction among the factors. RESULTS: PSO, UVA and PUVA all induced the apoptosis and the effects of PUVA were stronger than those of the other two. After HL-60 cells had been treated with PUVA, they all showed obvious ultrastructure changes due to apoptosis observed under the electron microscope. PSO, UVA and PUVA all decreased the expressions of FasL gene and protein. The effects of PUVA were stronger than those of the other two. CONCLUSIONS: PUVA can induce the apoptosis of HL-60 cells and the effects are stronger than those of PSO or UVA alone. The expression of FasL gene in HL-60 cells is down-regulated during the apoptosis induced by PUVA.

To study the Realgar induced T lymphocytic leukemia cell line CEM apoptosis in vitro.

Objective To establish a RP- HPLC method for the determination of naringin and neohesperidin in Fructus Aurantii. Methods The samples were extracted with methyl. The chromatographic conditions were as follows: Hypersil C18 Column( 250 mm? 4.6mm, 5 ? m) , mobile phase of acetonitrile- water(pH=3.0, volume ratio of 19 ∶ 81), detection wavelength at 283 nm, flow rate being 1.0 mL/min and the column temperature at 25 ℃ . Results The calibration curves were linear in the range of 0.156～ 0.934? g for narngin (r=0.9990) and 0.155～ 0.930 ? g for neohesperidin (r=0.9991) . The average recovery of naringin was 101.62 % (RSD=1.91 % ) , and that of nephesoeridin was 103.12 % (RSD=1.22 % ) . Conclusion This method is simple, accurate and reliable for the quality control of Fructus Aurantii.

Purpose:To study the expression of membrane protein HSP70 of K 562 /ADM cell using arsenic bisulfide(As 2S 2) treatment Methods:Membrane HSP70 and apoptosis of K 562 and multidrug resistant K 562 /ADM cell were studied by flow cytometry assay and RT PCR Results:Membrane HSP70 of K 562 and K 562 /ADM cell gradually increased from 12 to 24 hours with arsenic bisulfide treatment, and had higher expression at the level of HSP70mRNA Expression of membrane HSP70 and apoptosis of K 562 /ADM was higher than that of K 562 Conclusions:Arsenic bisulfide treatment can increase expression of membrane HSP70 of K 562 /ADM and K 562 cells.The change of membrane HSP70 is probably related to K 562 /ADM cell apoptosis

Objective To investigate the causes of thyroid carcinoma reoperation and to explore the tim-ing, surgical pattern of reoperation .Methods The clinicopathologic data of 127 patients undergoing reoperation for differentiated thyroid carcinoma ( DTC) from Sep 2006 to Sep 2011 were retrospectively analyzed .Results The residual rate of first thyroid operation was 54.3%, and the lymphatic metastasis rate was 31.5%.The rate of temporary and permanent recurrent laryngeal nerve injury after reoperation was 3.1%(4/127)and 1.6%(2/127) respectively .The rate of temporary and permanent hypoparathyroidism was 4.7%( 6/127 ) and 0.8%( 1/127 ) respectively .No recurrence of thyroid carcinoma or metastasis of cervical lymph node was reported after reopera -tion during a follow-up of 6 months to 6years.Conclusions Reoperation should be performed according to the initial operative and pathological findings in DTC patients who underwent local resection .The rate of complication in the reoperation , recurrence of thyroid carcinoma and metastasis of cervical lymph node can be effectively re -duced in meticulous and experienced hands .

Synthetic biology of traditional Chinese medicine (TCM) is a new and developing subject based on the research of secondary metabolite biosynthesis for nature products. The early development of synthetic biology focused on the screening and modification of parts or devices, and establishment of standardized device libraries. Panax notoginseng (Burk.) F.H.Chen is one of the most famous medicinal plants in Panax species. Triterpene saponins have important pharmacological activities in P. notoginseng. Squalene epoxidase (SE) has been considered as a key rate-limiting enzyme in biosynthetic pathways of triterpene saponins and phytosterols. SE acts as one of necessary devices for biosynthesis of triterpene saponins and phytosterols in vitro via synthetic biology approach. Here we cloned two genes encoding squalene epoxidase (PnSE1 and PnSE2) and analyzed the predict amino acid sequences by bioinformatic analysis. Further, we detected the gene expression profiling in different organs and the expression level of SEs in leaves elicited by methyl jasmonate (MeJA) treatment in 4-year-old P notoginseng using real-time quantitative PCR (real-time PCR). The study will provide a foundation for discovery and modification of devices in previous research by TCM synthetic biology. PnSE1 and PnSE2 encoded predicted proteins of 537 and 545 amino acids, respectively. Two amino acid sequences predicted from PnSEs shared strong similarity (79%), but were highly divergent in N-terminal regions (the first 70 amino acids). The genes expression profiling detected by real-time PCR, PnSE1 mRNA abundantly accumulated in all organs, especially in flower. PnSE2 was only weakly expressed and preferentially in flower. MeJA treatment enhanced the accumulation of PnSEI mRNA expression level in leaves, while there is no obvious enhancement of PnSE2 in same condition. Results indicated that the gene expressions of PnSE1 and PnSE2 were differently transcribed in four organs, and two PnSEs differently responded to MeJA stimuli. It was strongly suggested that PnSEs play different roles in secondary metabolite biosynthesis in P. notoginseng. PnSE1 might be involved in triterpenoid biosynthesis and PnSE2 might be involved in phytosterol biosynthesis.