Identification of chemical constituents in vitro and in vivo of Er Shen Zhenwu Decoction by utilizing ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry.

Er Shen Zhenwu Decoction is a prescription for treating chronic heart failure of heart and kidney yang deficiency, while its active ingredients remain unclear and difficult to identify. This paper aims to apply a rapid assay strategy of ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry to collect the mass spectrometry data of Er Shen Zhenwu Decoction and its decomposed recipes (monarch, minister, and assist). By comparing with retention time and MSE fragmentation patterns, 67 and 34 components in vitro and in vivo were identified, respectively, the main ingredients include saponins, terpenes, alkaloids, phenolic acids, tanshinone, urea, steroids, aromatics, organic acids, carbohydrates, and so forth, of which the monarch medicine > minister medicine > assist medicine. By comparison with reference standards, paeoniflorin, rosmarinic acid, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and atractylenolide III were identified in vitro and paeoniflorin, ginsenoside Rg1, ginsenoside Re and ginsenoside Rb1 were identified in vivo. In this study, the chemical ingredients of Er Shen Zhenwu Decoction were analyzed by ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry technology and each compound was grouped into the decomposed recipes. The identified substances can be used as references for Er Shen Zhenwu Decoction quality control and potential medicinal substances in chronic heart failure of heart and kidney yang deficiency treatment.

[The changes and significance of T lymphocytes and subsets before and after receiving highly active anti-retroviral therapy in children infected with HIV or AIDS patients].

OBJECTIVE: To investigate the changes of the T lymphocytes and their subsets before and after receiving highly active anti-retroviral therapy (HAART) in children who were infected with HIV or AIDS patients. METHODS: Ninety-nine children met the criteria were recruited. All of them had received HAART in Guangxi Center for Disease Control and Prevention from May 2006 to April 2009. Peripheral blood of 2 milliliter was collected before treatment (D0) and after 3, 6, 12, 18, 24, 30, and 36 months (M3, M6, M12, M18, M24, M30 and M36), respectively. Four-color fluorescence flow cytometry was used for the detection of the absolute numbers of CD3(+), CD4(+), CD8(+) T lymphocytes in peripheral blood. And then, the percentages of CD3(+), CD4(+), CD8(+) T lymphocytes in the CD45(+) cells and the ratio of CD4/CD8 were calculated. RESULTS: Sixteen-five (65.66%) cases were treated with lamivudine (3TC)/zidovudine (AZT)/nevirapine (NVP), and 16 (16.16%), 8 (8.08%) and 10 (10.10%) cases were treated with 3TC/stavudine (D4T)/NVP, 3TC/AZT/efavirenz (EFV) and 3TC/AZT/lpv-rtv (LPV/r), respectively. The median of the ratio of CD4/CD8 were 0.39, 0.51, 0.61, 0.65, 0.70, 0.73 and 0.76 in M3, M6, M12, M18, M24, M30 and M36, respectively which were significantly higher than that in D0 (0.19) (Z values were -5.158, -7.375, -9.078, -8.853, -8.373, -5.845 and -5.844 respectively, P < 0.000). The median of CD4% were 16.92%, 22.70%, 25.54%, 26.66%, 27.99%, 30.36% and 29.30% respectively in M3, M6, M12, M18, M24, M30 and M36 respectively, which were also higher significantly than that in D0 (9.92%) (Z values were -5.268, -7.568, -9.496, -9.171, -8.760, -6.190 and -5.964 respectively, P < 0.000). In addition, the median of the absolute numbers of CD4(+)T lymphocytes in peripheral blood were 631, 813, 1050, 946, 1057, 1166 and 894 cells/mm(3) respectively in M3, M6, M12, M18, M24, M30 and M36, which were higher significantly than that of D0 (382 cells/mm(3)) (Z values were -3.318, -5.288, -6.661, -5.886, -5.801, -4.110 and -3.600 respectively, P < 0.000). However, the median of CD8% were 47.25%, 43.01%, 43.04%, 42.60%, 41.37%, 40.83% and 38.31% respectively in M3, M6, M12, M18, M24, M30 and M36, which were lower significantly than that of D0 (53.17%) (Z values were -3.082, -4.697, -5.282, -5.846, -5.757, -3.883 and -4.380 respectively, P < 0.001). CONCLUSION: There is certain rules for the changes of T lymphocytes and their subsets, which may play important roles in the evaluation of the therapeutic effect and the clinical application guidance of HAART.