[Research of Yuanhu Zhitong prescription based on strategy of integrative pharmacology].

Chinese medicinal formulae (CMF) were usually used in the clinics of traditional Chinese medicines (TCM), which were critical for modernization of Chinese medicine to shed light on the interaction between CMF and biological organisms. However, correlation between system and part, macroscopic actions and microcosmic mechanism, ADME process and pharmacologic actions were usually neglected. The put-forward of integrative pharmacology provided a feasible approach to solve the problem of the fragmentation of TCM. For the past years, we applied the strategy of integrative pharmacology to study Yuanhu Zhitong prescription( YZP) systematically, and established two modes, chemical fingerprints-metabolism fingerprints-network targets and intestinal absorption-activity evaluation-data mining, to establish the interaction rule between the chemical composition and biological activity from multiple levels, such as the calculation and in vitro/vivo, which provided proof for the quality control, pharmacodynamic material basis and pharmacological action of YZP. In this paper, we summarized the related progresses of the research of YZP.

Tissue-specific distribution of ginsenosides in different aged ginseng and antioxidant activity of ginseng leaf.

The aim of this study was to systematically evaluate the effect of the cultivation year on the quality of different ginseng tissues. Qualitative and quantitative analyses of ginsenosides were conducted using a UPLC-UV-MS method. Eight main ginsenosides in three tissues (leaf, rhizome and main root) and four parts (periderm, phloem, cambium and xylem) of ginseng aged from 1 to 13 years were determined using a UPLC-PDA method. Additionally, the antioxidant capacities of ginseng leaves were analyzed by the DPPH, ABTS and HRSA methods. It was found that the contents of ginsenosides increased with cultivation years, causing a sequential content change of ginsenosides in an organ-specific manner: leaf > rhizome > main root. The ratio between protopanaxatriol (PPT, Rg1, Re and RF) and protopanaxadiol (PPD, Rb1, Rb2, RC and Rd) in the main root remained stable (about 1.0), while it increased in leaf from 1.37 to 3.14 and decreased in the rhizome from 0.99 to 0.72. The amount of ginsenosides accumulated in the periderm was 45.48 mg/g, which was more than twice as high compared with the other three parts. Furthermore, the antioxidant activities of ginseng leaves were measured as Trolox equivalents, showing that antioxidant activity increased along with time of cultivation. The results show that the best harvest time for shizhu ginseng is the fifth year of cultivation, and the root and rhizome could be used together within seven planting years for their similar PPT/PPD level. Besides, the quality of the ginseng products would be enhanced with the periderm. The ginseng leaf is rich in ginsenosides and has potential application for its antioxidant capacity.

[Effects of warm needling combined with zhangmo decoction on endometrial receptivity in patient with ovulation induction].

OBJECTIVE: To explore the effects of warm needling combined with Zhangmo decoction (see text) on endometrial receptivity in patients with clomiphene (CC)-induced ovulation. METHODS: One hundred and sixty cases were randomly divided into a CC group (group A), a CC+ progynova group (group B), a CC+ Zhangmo decoction group (group C) and a CC+ Zhangmo decoction + warm needling group (group D), 40 cases in each one. In the Group A, CC alone was applied. In the group B, progynova was jointly used on the 8th day of menstrual cycle. In the Group C, Zhangmo decoction was jointly used on the 5th day of menstrual cycle. In the group D, based on treatment of the Zhangmo decoction, warm needling was applied at Guanyuan (CV 4), Zhongji (CV 3) and Zigong (EX-CA 1) etc. The endometrial thickness and type, pulsatility index (PI), resistance index (RI), ratio of S/D on day of human chorionic gonadotropin (HCG) and pregnancy rate were observed in fou groups. RESULTS: The PI, RI and S/D in the group C and D were obviously lower than those in group A and B (all P < 0.01). The endometrial thickness was (7.7 +/- 1.49) mm in group B, (8.2 +/- 1.54) mm in group C and (8.9 +/- 1.51) mm in group D, which were significantly different from (6.4 5 +/- 1.26) mm in the group A (all P < 0.01) also there was a significant difference between group C and D (P < 0.05). The rate of endometrial type A was 65.0% in the group D, which was significantly higer than 27.5% in the group A, 32.5% in the group B and 35.0% in the group C (all P < 0.01). The pregnancy rate was 30.0% in the group D, which was obviously higher than 12.5% in the group A, 15.0% in the group B and 17.5% in the group C (P < 0.05). The endometrial thickness and rate of endometrial type A in the pregnant were obviously higher than those in the non-pregnant (both P < 0.01) while PI, RI and S/D was lower than those in the non-pregnant (P < 0.01, P < 0.05). CONCLUSION: Warm needing combined with Zhangmo decoction could improve endometrial thickness, morphology and uterine spiral artery to improve pregnancy rate, which has superior effect to clomiphene, clomiphene combined with progynova and clomiphene combined with Zhangmo decoction.

Screening of probiotic lactobacilli for inhibition of Shigella sonnei and the macromolecules involved in inhibition.

A total of 91 lactobacilli were screened for antimicrobial activity against Shigella sonnei. Agar-well assay showed that 16 lactobacilli displayed strong antibacterial activity against S. sonnei. The nature of these antimicrobial agents were investigated and shown to be dependent on their production of organic acids. Adhesion tests showed that 6 lactobacilli demonstrated good adherence to HT-29 cells, of these Lactobacillus johnsonii F0421 were selected for acid and bile salt tolerance properties. We further research on L. johnsonii F0421 inhibition of S. sonnei adhesion to HT-29 cells. The result showed that L. johnsonii F0421 exhibited significant inhibitory activity and excluded, competed and displaced adhered S. sonnei by 48%, 38% and 33%, respectively. In order to elucidate the inhibitory functions of macromolecules involved in L. johnsonii F0421, the cells were treated with 5 M LiCl, 0.05 M sodium metaperiodate and heating and assayed for inhibition activity. The results suggested a role of S-layer proteins on L. johnsonii F0421 cells in inhibition of the adhesion process, but carbohydrates do not seem to be involved. SDS-PAGE analysis confirmed the presence of S-layer proteins with dominant bands of approximately 40 kDa. In addition, 100 µg/well of S-layer proteins from L. johnsonii F0421 cells were effective in inhibiting adhesion of S. sonnei to HT-29 cells. These findings suggest that L. johnsonii F0421 possesses the capacity for inhibition of S. sonnei activity as well as probiotic properties, which could serve as a potential novel and effective probiotic strain for use in the food industry.

[Traditional medicine CDP and promoter demethylation in breast cancer cell lines].

OBJECTIVE: To investigate the demethylation effect of CDP on P16 and E-CADHERIN genes. METHODS: Breast cancer cell lines T47D and MDA-MB-435 were treated with CDP and DNA methyltransferase inhibitor 5-azacytidine (5-aza-C). The methylation of P16 and E-CADHERIN gene promoters were measured by methylation-specific PCR (MSP). The RNA transcription was determined by reverse transcription-PCR(RT-PCR). RESULTS: 1) The methylation-specific fragments of P16 gene promoter existed in T47D cells after 25, 50 and 75 micromol/L of CDP treatment for 6 days. An absolute demethylation on P16 gene occurred after treatment with 100 micromol/L of CDP. The unmethylation-specific fragments appeared in T47D cells after being treated with 25, 50, 75 and 100 micromol/L of CDP for 6 days. The RNA expression of P16 was detected after treatment with 75 and 100 micromol/L of CDP. 2) After being treated with 50 micromol/L of CDP, the methylation-specific fragments of CpG island in P16 gene promoter still existed in T47D cells. The unmethylation-specific fragments in T47D cells started to appear after 24 hours of treatment and lasted until 144 hour of treatment. The RNA expression was detected after 144 hours of treatment. 3) The demethylation on E-CADHERIN gene and genomic DNA or RNA transcription were not detected in MDA-MB-435 cells. CONCLUSION: CDP has concentration- and time-dependent demethylation effect on P16 gene in T47D cells, but not on E-CADHERIN gene in MDA-MB-435 cells, which indicates that CDP has substantial diversity in molecular activities.

[Isolation and identification of diterpenoids from Pinus koraiensis].

OBJECTIVE: To study chemical compounds from Pinus koraiensis. METHODS: The constituents were isolated by chromatographic method and the structures were identified on the basis of spectral alanlysis. RESULTS: Eight compounds were identified as 8 (14)-podocarpen-13-on-18-oic acid (1), 15-hydroxydehydroabietic acid (2), 12-hydroxyabietic acid (3), lambertianic acid (4), dehydroabietic acid (5), sandaracopimaric acid (6), beta-sitosterol (7), daucosterol (8). CONCLUSION: Compounds 1--6 are isolated from this plant for the first time.

[Influence of ganciclovir and astragalus membranaceus on proliferation of hematopoietic progenitor cells of cord blood after cytomegalovirus infection in vitro].

OBJECTIVE: Cytomegalovirus (CMV) infection was greatly common in the world. CMV infection produces usually mild or asymptomatic infections in individuals with normal immune responses, whereas it may cause serious disease in immunosuppressive patients. Clinical manifestations include suppression of myelopoiesis, a mononucleosis like syndrome, hepatosplenomegaly, lymphadenopathy, thrombocytopenia, and hemolytic anemia. In patients undergoing bone marrow transplantation CMV remains the most common infectious causes of morbidity and mortality. But the treatment drugs with specific effect for CMV was fewer at the present. This study was to investigate the effect of CMV on proliferation of colony forming unit granulocyte-macrophage (CFU-GM), CFU-erythroid (CFU-E), brust forming unit-erythroid (BFU-E), CFU-multipotential (CFU-Mix) and CFU-megakaryocyte (CFU-Mk) progenitor cells of cord blood (CB) with the presence of ganciclovir (GCV) and astragalus membranaceus in vitro. METHODS: Twenty CB samples were collected from fetal umbilical vein of normal term spontaneous delivery neonates. The colony forming unit-assay was applied to observe the suppression effect of CMV-AD169 strain on CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk of CB with the presence of GCV and astragalus membranaceus in vitro. The technique of PCR was used to demonstrate the existence of CMV-AD169 DNA in the colony cells of cultured CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk. RESULTS: (1) The numbers of CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk colonies in CMV infection groups were significantly less than those in blank and mock group, respectively. The last time of colonies in groups with CMV infection was significantly shorten compared with the blank and mock group. (2) CMV-DNA was positively detected in the colony cells of CMV infection groups by PCR, while negative in the control groups. (3) The lasting time of CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk colonies infected with CMV extended significantly with the presence of astragalus membranaceus and GCV, and the numbers of those increased significantly compared with the CMV infection group, respectively. The increasing rate of colonies was 27.2%, 45.2%, 49.1%, 39.0% and 11.9% with astragalus membranaceus group, 37.4%, 74.2%, 71.7%, 67.4% and 38.9% with GCV group, 53.6%, 83.8%, 88.7%, 87.8% and 61.5% with astragalus membranaceus and GCV group, respectively. CONCLUSIONS: The differentiation and proliferation of CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk were significantly inhibited after infected with CMV-AD169 strain. The suppression effect of CMV-AD169 on CFU-GM, CFU-E, BFU-E, CFU-Mix and CFU-Mk was inhibited with the presence of GCV and astragalus membranaceus in vitro. This suggested that CMV-AD169 may be inhibited or killed by GCV and Astragalus Membranaceus in vitro.