Phloroglucinol Derivatives from the Fruits of Eucalyptus globulus and Their Cytotoxic Activities.

A new phloroglucinol derivative, named eucalyptin B (1), along with five related known compounds (2 - 6), was isolated from the fruits of Eucalyptus globulus. Their structures were elucidated by means of 1D- and 2D-NMR spectroscopy, with the absolute configuration of 1 determined by electronic circular dichroism (ECD) calculations. All isolated compounds (1 - 6) were evaluated for their cytotoxic activities against lung (A549), breast (4T1), and skin (B16F10) cancer cell lines. On the basis of cell viability assay, the cytotoxic activity of eucalyptin B (1) was further confirmed by apoptosis assay. Additionally, after treatment with eucalyptin B (1), the apoptosis factor proteins (Bcl2 and Bax) and caspase-3 levels in A549 cells were also determined by Western-blot analysis. By cytotoxic assay, eucalyptin B (1) exhibited potent cytotoxicity against A549 cells with an IC50 value of 1.51 µm and induced concentration dependent apoptosis of up to 49%. Additionally, eucalyptin B (1) inhibited 5-fold and increased 10-folds in the level of Bcl2 and Bax, respectively. Furthermore, the 11-fold increase in the level of caspase-3 confirmed eucalyptin B (1) activated caspase dependent apoptosis pathway. In conclusion, the isolated compound eucalyptin B (1) has promising cytotoxic activity in tumor cells, especially in A549.

[Study on macrophages infected with Mycobacterium tuberculosis H37Ra in vitro].

AIM: To study the production of nitric oxide and secretion of cytokines after infection of macrophages with Mycobacterium tuberculosis H37Ra. METHODS: 24 hours after infection of RAW264.7 cells with Mycobacterium tuberculosis H37Ra, the production of NO and H(2);O(2); as well as the secretion levels of IL-12 and TNF-α in the supernatants of culture were determined by Griess method, chemical method and ELISA assay respectively. The expression of IL-12 and TNF-α mRNA in macrophages was detected by reverse transcription polymerase chain reaction (RT- PCR). RESULTS: Macrophages infected with Mycobacterium tuberculosis H37Ra produced effectively more NO, H(2);O(2);, and enhanced the release of IL-12, TNF-α and the expression of IL-12, TNF-α mRNA (P<0.05). CONCLUSION: Mycobacterium tuberculosis H37Ra can induce the production of more nitric oxide and cytokines which play important roles in the host immune response.

[The influence of benign prostatic hyperplasia drugs on incidence and pathology grading of prostate cancer].

OBJECTIVE: To analyze the influence of benign prostatic hyperplasia (BPH) drugs on incidence and pathology grading of prostate cancer in China. METHODS: Retrospectively investigated the history of drug treatment in 1029 cases of BPH in patients from February 1998 to December 2004. According to the history of drug use, the patients were divided into 4 groups: finasteride group, alpha-receptor inhibitor group, finasteride and alpha-receptor inhibitor combination group and control group (untreated group). We gathered pathology sections of patients in all groups, and gave Gleason Score to each. The difference of incidence and pathology grading of prostate cancer were analyzed by Stata 7.0. RESULTS: The incidence of prostate cancer in the population of our study was 13.5%; The incidence in finasteride group, alpha-receptor inhibitor group, combination group and control group was 9.8%, 16.0%, 10.3% and 18.6%, respectively. There was significant difference between the two groups with the use of finasteride and the two groups without it (P < 0.05). In our study, the ratio of middle or high level pathology grading (Gleason ≥ 7) in prostate cancer patients was 58.3%, the ratio of middle or high level pathology grading prostate cancer patients in the four groups was 71.4%, 59.6%, 67.7% and 40.0%, respectively. In the comparison of composition ratio of middle or high level prostate cancer, there was significant difference between the two groups with the use of finasteride and the two groups without it (P < 0.05). CONCLUSIONS: Finasteride can lower the risk of prostate cancer, but increase the pathology grade of the prostate cancer which has occurred in the same time. The alpha-receptor inhibitor does not have the same effect.

[Study on the cytotoxicity of spleen lymphocytes and immune mechanisms in mice immunized by Mycobacterium tuberculosis H37Ra].

AIM: To explore the specific cytotoxicity of spleen lymphocytes and the immune mechanisms in mice immunized by Mycobacterium tuberculosis(MTB) H37Ra. METHODS: At the various interval (30 days, 60 days) after the mice were immunized by MTB H37Ra, BCG and PBS, the spleen lymphocytes of the immunized mice were used as the effector cells while the Sp2/0 cells expressing the secreted protein Ag85B were used as the target cells, and the cytotoxicity of spleen lymphocytes in the immunized mice was measured by MTT assay. Spleen lymphocytes were collected at 60 days after immunization and stimulated with PPD, and the expression of perforin, granzyme B on mRNA level was detected by RT-PCR. RESULTS: The cytotoxicity of spleen lymphocytes in the group immunized by MTB H37Ra was significantly higher than that of PBS control group (P<0.05), and slightly higher than that of BCG group. The mRNA expression of perforin, granzyme B in H37Ra group and BCG group was significantly higher than that in PBS control group (P<0.05); the expression of perforin mRNA in H37Ra group was significantly higher than that in BCG group (P<0.05), but there was no obvious difference with regard to the expression of granzyme B mRNA between H37Ra group and BCG group. CONCLUSION: The cytotoxicity of spleen lymphocytes is enhanced after mice were immunized by MTB H37Ra, which may be related to the expression of perforin and granzyme B.

Memory performance of hypercholesterolemic mice in response to treatment with soy isoflavones.

The aim of this study is to investigate the memory performance of hypercholesterolemic mice in response to soy isoflavones (SI) treatment and the mechanism involved. In this study, 64 mice were randomly divided into four groups: control, high lipid diet without SI, high lipid diet with a low SI level (50 mg/kg bw) and high lipid diet with a high SI level (100 mg/kg bw). The experimental period was 30 days. The results indicated that the mice given the different treatments showed the different percentages of good, medium and poor memory performance. chi(2) analysis revealed significant difference in memory performance (P<0.05) between the high lipid diet without SI group and the high lipid diet with a low SI level group or high lipid diet with a high SI level group. Moreover, SI treatment resulted in a decrease in blood cholesterol (TC) level (high lipid diet without SI group versus high lipid diet with a low SI level group or high lipid diet with a high SI level group, P<0.05) and triglyceride (TG) level (high lipid diet without SI group versus high lipid diet with a low SI level group or high lipid diet with a high SI level group, P<0.05). In addition, SI treatment resulted in a significant decrease in acetylcholinesterase (AChE) activity and significant increases in glutamic acid and aspartic acid contents in the frontal cerebral cortex and hippocampus. The results suggest that SI improve the memory performance of hypercholesterolemic mice, and the mechanism underlying the improvement might closely correlate with its roles in decreasing high blood lipid levels and modulating the metabolism of neurotransmitters such as acetylcholine and amino acids in brain areas of hypercholesterolemic mice.

[Advances in the chiral resolution of organophosphorus compounds by liquid chromatography].

The developments in the chiral resolution of organophosphorus compounds by liquid chromatography in recent years are reviewed. Two different approaches of separation: indirect resolution (chiral derivatization reagent method) and direct resolution (chiral mobile phase additive and chiral stationary phase), focusing on several chiral stationary phases--Pirkle type, cyclodextrin, cellulose, ligand exchange and other chiral stationary phases are presented. The possible mechanism of chiral recognition is discussed. Eighty-five references are involved.