Direct infusion electrospray ionization-ion mobility-mass spectrometry for rapid metabolite marker discovery of medicinal Phellodendron Bark.

Ion-mobility mass spectrometry (IM-MS) currently serves as a powerful tool for the structural identification of numerous biological compounds and small molecules. In this work, rapid metabolomic analysis of closely-related herbal medicines by direct injection ion mobility-quadrupole time-of-flight mass spectrometry (DI-IM-QTOF MS) was established. Phellodendron chinense Bark (PC) and Phellodendron amurense Bark (PA) were studied as a case. Thirty-three batches of PC and twenty-two batches of PA have been directly injected in electrospray ionization-IM-QTOF MS in positive mode. Without chromatographic separation, each run was completed within 3 min. After data alignment and statistical analysis, a total of seven chemical markers were found (p-value < 0.05, VIP > 1.00). Among them, the ion m/z 342.17 and m/z 356.18 present a single peak in the drift spectrum, respectively, but their drift time has a certain deviation compared with the pure substance of known compounds. In addition, the MS/MS spectra also confirmed that the single peak includes two chemical isomers. To investigate the composition ratio of individual isomers, the calibration curves of relative drift time (rDT) based on the standard superposition method were established, which were found to fit the least square regression. The ion [M]+m/z 342.17 was recognized consisting of magnoflorine (MAG) and phellodendrine (PHE), and their composition ratio in PA and PC samples was calculated. The results were compared with those obtained by the HPLC quantitative method, which produced equivalent quantification results. Our DI-IM-QTOF MS methodology provides an additional methodology for the relative quantification of unresolved isomers in drift tube IM-MS and offers DI-IM-QTOF MS based metabolomics.

A multicenter randomized open-label study of rituximab plus rhTPO vs rituximab in corticosteroid-resistant or relapsed ITP.

This study aimed to compare the efficacy and safety of rituximab (RTX) plus recombinant human thrombopoietin (rhTPO) with RTX alone in patients with immune thrombocytopenia (ITP) who had failed to respond to corticosteroids or relapsed. Recruited patients were randomized at a ratio of 2:1 into 2 groups: the combination group (RTX + rhTPO, n = 77) and the monotherapy group (RTX, n = 38). Overall response was achieved in 79.2% of patients in the combination group vs 71.1% in the monotherapy group (P = .36), and the complete response (CR) rate was 45.4% in the combination group compared with 23.7% in the monotherapy group (P = .026). The combination group had significantly shorter time to response (TTR; median and range, 7 and 4-28 days) compared with the monotherapy group (28 and 4-90 days) (P < .01). There was no difference between these 2 groups in terms of the long-term response (P = .12). Our findings demonstrated that the combination of RTX and rhTPO significantly increased the CR rate and shortened TTR compared with RTX monotherapy in the treatment of corticosteroid-resistant or relapsed ITP but failed to show a beneficial effect on the long-lasting response. This study is registered at www.clinicaltrials.gov as #NCT01525836.

[Study of WIF-1 promoter methylation with expressions of ß-catenin in acute leukemia].

OBJECTIVE: To explore the significance of WIF-1 gene promoter methylation and the expression of ß-catenin in acute leukemia (AL) patients. METHODS: The method of methylation specific polymerase chain reaction was employed to detect the status of WIF-1 gene methylation in 55 acute leukemia patients and normal controls from January 2009 to June 2010 in our hospital. The expression of ß-catenin was measured by flow cytometry. RESULTS: The methylation of WIF-1 gene promoter was found in 32.7% (18/55) AL patients. And the percentage was significantly higher than that of the controls (0). The patients with the methylation of WIF-1 gene had a lower complete remission rate (38.9%, 7/18) for the first chemotherapy than those without (81.1%, 30/37) (P < 0.05). The expressions of ß-catenin in methylation AL patients and those with non-methylation were 17.5% ± 3.3% and 15.4% ± 3.6% respectively. And they were significantly higher than the controls (10.5% ± 1.5%, P < 0.05). The expression of ß-catenin was higher in positive methylation patients than those negative ones (P < 0.05). CONCLUSION: The methylation of WIF-1 gene promoter and ß-catenin may be involved in the abnormal activation of Wnt/ß-catenin signal in acute leukemia.

[A preliminary study on the implication of Apaf-1 promoter methylation and the expression of apoptosis inhibitor protein Apollon in adult acute leukemia].

OBJECTIVE: To investigate the role of Apaf-1 gene promoter methylation and apoptosis inhibitor protein Apollon in pathogenesis of acute leukemia (AL) and their clinical significance. METHODS: Methylation specific PCR (MSP) was used to detect the methylation status of Apaf-1 gene promoter in 53 AL patients (28 AML, 10 ALL and 15 relapsed) and 10 healthy or nonmalignant blood diseases patients as control. RT-PCR was used to detect the expression levels of Apaf-1 mRNA and immunocytochemistry to detect the expression levels of Apollon protein. RESULTS: The abnromal methylation of Apaf-1 gene promotor in AL was 18/53(33.9%). No Apaf-1 mRNA was detected in methylation positive patients. Only one case in healthy and nonmalignant individuals was deletion of Apaf-1 mRNA expression without abnormal methylation. The positive methylation rate in AL bone marrow mononuclear cells was significantly higher than that in controls (P < 0.05). The expressin levels of Apollon protein in AL patients was higher than that in control (P < 0.05). The positive methylation ratio and Apollon protein level were higher in white blood cell count > 10 × 10(9)/L than in ≤ 10 × 10(9)/L (P < 0.05). There is a positive correlaiton between positive methylation ratio and Apollon protein expression in AL patients. CONCLUSION: Abnormal methylation of Apaf-1 gene promotor and high expression of Apollon might involved in leukemogenesis.