Humulus lupulus L. Extract Prevents Ovariectomy-Induced Osteoporosis in Mice and Regulates Activities of Osteoblasts and Osteoclasts / 中国结合医学杂志

OBJECTIVE@#To systematically evaluate the protective effects of Humulus lupulus L. extract (HLE) on osteoporosis mice.@*METHODS@#In vivo experiment, a total of 35 12-week-old female ICR mice were equally divided into 5 groups: the sham control group (sham); the ovariectomy with vehicle group (OVX); the OVX with estradiol valerate [EV, 0.2 mg/(kg•d)] the OVX with low- or high-dose HLE groups [HLE, 1 g/(kg•d) and 3 g/(kg•d)], 7 in each group. Treatment began 1 week after the ovariectomized surgery and lasted for 12 weeks. Bone mass and trabecular bone mircoarchitecture were evaluated by micro computed tomography, and bone turnover markers in serum were evaluated using enzyme-linked immunosorbent assay (ELISA) kits. In vitro experiment, osteoblasts and osteoclasts were treated with HLE at doses of 0, 4, 20 and 100 µg/mL. Biomarkers for bone formation in osteoblasts and bone resorption in osteoclasts were analyzed.@*RESULTS@#Compared with the OVX group, HLE exerted bone protective effects by the increase of estradiol (P<0.05), the improvement of cancellous bone structure, bone mineral density (P<0.01) and the reduction of serum alkaline phosphatase (ALP), tartrate resistant acid phosphatase (TRAP), bone gla-protein, c-terminal telopeptides of type I collagen (CTX-I) and deoxypyridinoline levels (P<0.01 for all). In vitro experiment, compared with the control group, HLE at 20 µg/mL promoted the cell proliferation (P<0.01), and increased the expression of bone morphogenetic protein-2 and osteopontin levels in osteoblasts (both P<0.05). HLE at 100 µg/mL increased the osteoblastic ALP activities, and HLE at all dose enhanced the extracellular matrix mineralization (both P<0.01). Furthermore, compared with the control group, HLE at 20 µg/mL and 100 µg/mL inhibited osteoclastic TRAP activity (P<0.01), and reduced the expression of matrix metalloproteinase-9 and cathepsin K (both P<0.05).@*CONCLUSION@#HLE may protect against bone loss, and have potentials in the treatment of osteoporosis.

Study on exosite inhibitor of cathepsin K from Herba Ecliptae fractions / 药学学报

Cathepsin K (CTSK) is considered a critical pharmaceutical target in the treatment of osteoporosis. CTSK exerts proteolytic activities against regulatory proteins besides its collagenase function, which may account for some of the adverse reactions when blocked by active site-directed inhibitors. Exosite inhibitors that can discriminate between the therapeutic collagenase and other biological activities of CTSK specifically inhibit the collagenase activity of CTSK without interfering with the other proteolytic activities of the protease. Active recombinant CTSK was expressed in Pichia pastoris, and purified by n-butyl sepharose and SP sepharose column chromatography. Herba Ecliptae is a common traditional Chinese medicine in the treatment of bone diseases. Collagenase assay and benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin (Z-FR-MCA) substrate assay based on CTSK are applied to verify the exosite inhibitors. n-Butanol extract of Herba Ecliptae are the most active fraction and eclalbasaponin IX isolated from n-butanol fraction is the potential exosite inhibitor of CTSK.

Research progress of chemical constituents and pharmacological activities in Humulus lupulus / 中国中药杂志

Hops, the female inflorescences of the hop plant (Humulus lupulus), are widely used in the brewing industry to add bitterness and aroma to beer. Combining with the relevant literature, the chemical composition(resinae, volatile oil, polyphenol and polysaccharide) in hops and their pharmacological effects are reviewed in this paper so as to present some sights for further application research and development.

Dynamic changes of flavonoids in Actinidia valvata leaves at different growing stages measured by HPLC-MS/MS / 中国天然药物

Flavonoids are a large group of phenolic secondary metabolites havinga wide range of biochemical and pharmacological effects. Quantitative analysis of flavonoid profiles in the genus Actinidia, which has not been intensively conducted, is useful to a better understanding of the pattern and distribution of flavonoids. In the present work, a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method was developed to profile the flavonoids, which was then used to determine the dynamic change of 17 biologically active flavonoids in the leaves of Actinidia valvata at the main growing stages, including glucuronides and acylated di- and triglycosides of flavonoids. The contents of flavonoid triglycosides were significantly higher than other flavonoids. The highest concentrations of kaemperol glycosides were observed in June, while other flavonoids showed highest concentrations in October. On the other hand, the contents of four isorhamnetin glycosides were increased sharply in September to October. The flavonoid profiles seem to be related to temperature, UV-B, and water deficit. Further studies are required to examine the functions of flavonoids in the Actinidia valvata and the underlying molecular mechanisms of actions.

Antipyretic Effects of Eupatorium chinense and Its Mechanism / 中草药·英文版

Objective: To investigate the antipyretic effect of Eupatorium chinense and its mechanism. Methods: The content of arginine vasopressin (AVP) in ventral septal area (VSA) and blood plasma, and cyclic adenosine monophosphate (cAMP) levels of hypothalamus and blood plasma were determined by enzyme linked immunosorbent assay. Results: The body temperature (Tb) was decreased at 1 h after administration of E. chinense (3 and 6 g/kg) and Aspirin (0.3 g/kg) respectively, which was significantly different from the temperature of fever model group. The antipyretic effect of Aspirin and E. chinense lasted for longer time. Aspirin (0.3 g/kg) and E. chinense (3 and 6 g/kg) reduced the level of cAMP in hypothalamus of fever rats and increased AVP content in plasma. The changes in cAMP content in plasma of all drug treatment groups were not obvious. Conclusion: E. chinense has strong antipyretic effect and may affect the production of AVP and cAMP in fever rats.

Research on immune-modulation effects and its mechanisms of Guizhi Fuling capsule and its active ingredient combination / 中国中药杂志

The effects of Guizhi Fuling capsule and its active ingredient combination within different concentration on SPL proliferate were observed by MTT method. The ratio of CD80/86, CD3CD25 and CD3CD69 was used to evaluate cell activation effects of Guizhi Fuling capsule and its active ingredient combination by FCM. Guizhi Fuling capsule with concentration of 400 mg · L(-1)can promote spleen lymphocyte proliferation, as well as the active ingredient combination, which showed the obvious dose-effect relationship. Compared with control group, the difference has statistical significance (P≤0.01). The result of FCM showed that Guizhi Fuling capsule and its active ingredient combination can promote CD80 and CD86 expression on spleen lymphocyte, and also can increase CD25 and CD69 ratio between spleen CD3+ cells. Compared with control group, the difference has statistical significance (P≤0.01). Thus, Guizhi Fuling capsule and its active ingredient combination may have immune-modulate effects, and the mechanism may have a close relationship with the lymphocyte activation.

Transient expression of exogenous human platelet-derived growth factor-B in gingival fibroblasts of dog / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore transient expression of the eukaryotic expression plasmid carrying human platelet-derived growth factor-B (hPDGF-B) in gingival fibroblasts of Beagle dog.</p><p><b>METHODS</b>Plasmid carrying hPDGF-B (EX-A0380-M03) was amplified and identified, and then transfected into gingival fibroblasts of Beagle dog. Reverse transcription polymerase chain reaction (RT-PCR), immunocytochemistry, enzyme-linked immunosorbent assay (ELISA) and Western bolt were choose to detect the expression of hPDGF-B.</p><p><b>RESULTS</b>Target gene carried by EX-A0380-M03 was hPDGF-B. Green fluorescence protein (GFP) expressed by transfected gingival fibroblasts was observed under inverted phase contrast fluorescence microscope (IPCFM) (after 24 hours) and the transfection efficiency was 18%-38% (after 48 hours). Serials other methods (RT-PCR, immunocytochemistry, and ELISA) mentioned above also convinced that cells expressed hPDGF-B, and the protein that was a kind of fusion protein composed of PDGF-BB and GFP was identified by Western blot.</p><p><b>CONCLUSION</b>Eukaryotic expression plasmid carrying hPDGF-B was transfected into gingival fibroblasts successfully, and a kind of fusion protein was expressed.</p>

Protein expression by planktonic and biofilm cells of clinical isolations of Streptococcus mutans / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the changes in the surface-associated protein expression of planktonic cells and biofilm cells of clinical isolations of Streptococcus mutans (Sm).</p><p><b>METHODS</b>The proteins were extracted by the method of Homer from the nonadhered planktonic and the adhered biofilm cells and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis followed by image analysis. Proteins were identified by matrix-assisted laser desorption time of flight mass spectrometry and computer-assisted protein sequence analysis.</p><p><b>RESULTS</b>Image analysis revealed that 30% - 31% of the protein spots in biofilm was modulated. A total of 238 proteins changed 1.3-fold or greater in biofilm cells compared to planktonic cells in Sm18, with 5 only expressed in biofilm cells and 5 not expressed in biofilm cells. A total of 279 proteins changed 1.3-fold or greater in biofilm cells compared to planktonic cells in Sm593, with 12 only expressed in biofilm cells and 2 not expressed in biofilm cells. Two clinical isolations in biofilm cells had three identical protein expressions whose functions were associated with biosynthetic processes.</p><p><b>CONCLUSIONS</b>The two clinical isolations in biofilm status have high expression of some special proteins, which are presumed to be key proteins essential for formation of biofilm. The difference of protein expression in biofilm cells of the two clinical isolations may represent their distinct characters.</p>

Functional changes of dendritic cells after infection by recombinant retrovirus carrying human telomerase reverse transcriptase gene fragment / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the functional changes of dendritic cells (DCs) after infection by recombinant retrovirus carrying human telomerase reverse transcriptase (hTERT) gene fragment.</p><p><b>METHODS</b>Interleukin-12 (IL-12) levels in DC culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA). The abilities of DCs infected with recombinant retrovirus carrying hTERT gene (hTERT-DCs) and non-infected DCs (N-DCs) to stimulate allogeneic lymphocyte proliferation were evaluated with mixed leukocytes reaction (MLR), and the surface markers of DCs including CD80, CD83, CD86 and HLA-DR were detected by flow cytometry. Cytotoxic T lymphocyte (CTL) assay was performed with CytoTox 96 non-radioactive cytoxicity assay.</p><p><b>RESULTS</b>Compared with N-DCs, hTERT-DCs showed no significant changes in IL-12 secretion and capacity to stimulate allogeneic lymphocytes reaction, but had significantly lower CD83 expression. Specific CTLs induced by hTERT-DCs resulted in higher cytotoxicity against telomerase-positive target cells than that against the negative target cells.</p><p><b>CONCLUSION</b>Infection with the recombinant retrovirus carrying hTERT fragment may jeopardize the maturation of DCs, which, however, still retain their capacity to activate and stimulate lymphocyte proliferation and to prime autologous T lymphocytes to generate specific CTL against hTERT.</p>

Protective effect of curcumin on experimental liver injury in mice / 中国中药杂志

<p><b>OBJECTIVE</b>To Study the protective effect of curcumin on three models of experimental liver injury in mice.</p><p><b>METHOD</b>The experimental models of live injury were induced by carbon tetrachloride (CCl4), D-galactosamine (D-Gal N), and Bacillus Calmette-Guerin (BCG) Plus lipolysaccharides (LPS), respectively, in mice. The serum ALT, AST, NO and liver MDA were measured to evaluate the protective effect of curcumin on experimental injury in mice.</p><p><b>RESULT</b>Curcumin (50 mg.kg-1, 100 mg.kg-1, 150 mg.kg-1), like biophenyldicarboxylate, were shown to significantly inhibit the increase of serum ALT, AST, NO and liver molondialdehyde (MDA) content induced by CCl4, D-Gal N, BCG + LPS.</p><p><b>CONCLUSION</b>Curcumin showed protective effect against liver injury induced by CCl4, D-Gal N, BCG plus LPS.</p>