Three new cytotoxic oleanane triterpenoid saponins from Cylindrokelupha dalatensis.

Three new minor oleanane triterpenoid saponins, cylindrosides B (1), C (2), and D (3), were isolated from the seed of Cylindrokelupha dalatensis using chromatographic method. Their structures were established on the basis of the chemical and spectroscopic evidences. They displayed significant antitumor activity in vitro against HL60 cancer cell lines and IC50 values were 7.15 ± 0.63, 10.07 ± 0.97, and 4.74 ± 0.57 µM, respectively, by MTT method.

[Effect of Aralia echinocaulis containing serum on Wnt/ß-catenin signaling pathway of primary osteoblast].

This paper was aimed to investigate the effect of Aralia echinocaulis containing serum on expression of ß-catenin, Wnt-1, Frizzed-2, TCF and Axin in Wnt/ß-catenin signaling pathway of primary osteoblasts. SD healthy female rats (n=80) were used to make A. echinocaulis containing serum by gastric perfusion for seven days with distilled water, A. echinocaulis decoction high dosage, middle dosage, and low dosage. In vitro, primary osteoblasts were cultured and identified. The third generation primary osteoblasts were taken and cultured for 48 h, then cells were treated with the different drug serums for 10 days and calcified nodules were counted by alizarin red staining. The cells were collected after treatment for 48 h and the expression levels of ß-catenin, Wnt-1, Frizzled-2, TCF and Axin were detected by Real-time PCR and Western blot. The results suggested that the in vitro cells were primary osteoblasts; and after treatment, various doses groups could promote the mineralization ability of primary osteoblasts, up-regulate the mRNA and protein expression levels of ß-catenin, Wnt-1, Frizzled-2, and TCF, and down-regulate the mRNA and protein expression levels of Axin. These findings indicated that A. echinocaulis containing serum can enhance the differentiation and proliferation of osteoblasts by regulating the expression levels of ß-catenin, Wnt-1, Frizzled-2, TCF and Axin in Wnt/ß-catenin signaling pathway of primary osteoblasts.

Dynamic changes in the migratory microbial components of colon tissue during different periods of sepsis in an LPS-induced rat model.

Previous studies have shown that bacterial translocation may play an important role in worsening gastrointestinal injury during sepsis. However, the dynamics of specific microbiota components in intestinal tissues at different sepsis stages remain unclear. Rats receiving intraperitoneal lipopolysaccharide (LPS) were sacrificed at 12 h and 48 h post-injection. Routine blood, serum cytokines, and microbiota in colon tissue, colonic contents, and lung tissue at different time points were assessed. Migratory microbial components in colonic tissue at 12 h and 48 h post-LPS were identified using source tracking, characteristic component identification, and abundance difference analyses. Colonic tissue microbiota changed dynamically over time after LPS injection, involving translocation of microbial components from colon contents and lung tissue at different time points. Bacteria migrating to colon tissue at 12 h sepsis were mainly from colonic contents, while those at 48 h were predominantly from the lung tissue. The migratory microbial components in colon tissue were widely associated with blood indicators and colonizing genus abundance and microbiota functionality in colon tissue. In this study, the temporal dynamics of bacterial translocation from various sources into colon tissues at different sepsis progression stages were characterized for the first time, and the species composition of these migrating microbes was delineated. These bacterial migrants may contribute to the pathophysiological processes in sepsis through direct interactions or indirectly by modulating colonic microbiota community structure and function.

Proteasome inhibition-enhanced fracture repair is associated with increased mesenchymal progenitor cells in mice.

The ubiquitin/proteasome system controls the stability of Runx2 and JunB, proteins essential for differentiation of mesenchymal progenitor/stem cells (MPCs) to osteoblasts. Local administration of proteasome inhibitor enhances bone fracture healing by accelerating endochondral ossification. However, if a short-term administration of proteasome inhibitor enhances fracture repair and potential mechanisms involved have yet to be exploited. We hypothesize that injury activates the ubiquitin/proteasome system in callus, leading to elevated protein ubiquitination and degradation, decreased MPCs, and impaired fracture healing, which can be prevented by a short-term of proteasome inhibition. We used a tibial fracture model in Nestin-GFP reporter mice, in which a subgroup of MPCs are labeled by Nestin-GFP, to test our hypothesis. We found increased expression of ubiquitin E3 ligases and ubiquitinated proteins in callus tissues at the early phase of fracture repair. Proteasome inhibitor Bortezomib, given soon after fracture, enhanced fracture repair, which is accompanied by increased callus Nestin-GFP+ cells and their proliferation, and the expression of osteoblast-associated genes and Runx2 and JunB proteins. Thus, early treatment of fractures with Bortezomib could enhance the fracture repair by increasing the number and proliferation of MPCs.

Three new triterpenoid saponins from Aralia echinocaulis.

Objective: To study the active ingredients in the root bark of Aralia echinocaulis. Methods: Three triterpenoid saponins were separated from the 70% ethanol extracts and purified by column chromatography and their structures were determined by spectroscopic analysis. Compound 1 and 3 were evaluated for antioxidant activity by the in vitro DPPH free radical scavenging ability and the protective effect of OH- induced DNA oxidative damage. Results: Compound 1 was a new type of triterpenoid saponin, named as echinocaulisaglycone 3-O-ß-D-glucopyranoside (echinocaulisaponin A), and it had good antioxidant activity. Compound 2 was similar to compound 1, named as 1-hydroxyl-echinocaulisaglycone 3-O-ß-D-glucopyranoside (echinocaulisaponin B). Compound 3 was also a new type of triterpenoid saponin, named as echinocaulisaglycone II 3-O-α-L-arabinopyranosyl-(1″→4')-ß-D-glucopyranosiduronic acid (echinocaulisaponin C), and its antioxidant activity was weaker than compound 1. Conclusion: In this study, three new compounds were discovered and two of them were carried out in vitro anti-oxidation studies, laying the foundation for further research on the treatment of related diseases (cardiovascular disease, arthritis, age-related macular degeneration, etc.) through anti-oxidation or quenching free radical function.

Dynamic alterations in the lung microbiota in a rat model of lipopolysaccharide-induced acute lung injury.

The lung microbiota have been found to be substantially altered in numerous pulmonary disorders, and crosstalk between the host pathophysiology and lung microbiota plays critical roles in the regulation of disease states. The aim of this study was to investigate dynamic changes in the lung microbiota during different stages of acute lung injury and acute respiratory distress syndrome (ALI/ARDS). Rats receiving an intraperitoneal administration of lipopolysaccharide (LPS) were sacrificed at 12 and 48 h after injection, and the hematological parameters, serum cytokine levels, and histological characteristics of the lung tissue and lung microbiota were assessed. After LPS injection, along with fluctuations of systemic cytokine levels and the onset and regression of pulmonary edema, the diversity, components, and functionalities of the pulmonary microbiota underwent significant dynamic changes. The volatility of the α-diversity indices narrowed after LPS injection, and the indices significantly decreased 48 h later. The abundance of 18 genera and functionality of adenosine triphosphate-binding cassette (ABC) transporters, pentose phosphate, and bacterial chemotaxis pathways were found to significantly differ between specified time points. Several significant correlations between the components and functionalities of the lung microbiota and indicative symptoms of ALI/ARDS were also observed. Brevibacterium was correlated with cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-10, and IL-6 and with hematological percentage of neutrophils (NEU%); Wnt, Notch, and chronic myeloid leukemia signaling pathways were correlated with IL-1ß; mitogen-activated protein kinase (MAPK) signaling pathway-yeast was correlated with IL-10; and the pathways of ascorbate and aldarate metabolism and basal transcription factors were correlated with platelet-related indicators. The correlations between the lung microbiota and indicative symptoms of ALI/ARDS identified in this study support further investigation into the underlying mechanism of host-microbiota interactions during lung injury and repair.

[Applications and approved projects on traditional Chinese medicine in National Natural Science Foundation of China in 2010].

In this article, the authors firstly summarized the number of applications submitted to and projects supported by the National Natural Science Foundation of China (NSFC) in the field of traditional Chinese medicine research in 2010. Then they described the district distribution, research direction layout and allotment of the approved projects in the three primary disciplines (traditional Chinese medicine, Chinese materia medica and integrated traditional Chinese and Western medicine) and their 43 subdisciplines. The targeting suggestions for improvement were given respectively by concluding the reason of disapproved projects from the point of view of applicants and supporting institution, and by stating the common problems existing in the review process from the perspectives of fund managers and evaluation experts. Lastly, the major funding fields in the near future were predicted in the hope of providing guidance for applicants.

[Determination of eight metal elements in Cichorium glandulosum Boiss et Huet by microwave digestion-FAAS].

The Cichorium glandulosum Boiss et Huet is a traditional Uighur natural herbal medicine, but has not been analyzed and studied in terms of its metal elements. In the experiment, the Cichorium glandulosum Boiss et Huet powder was digested with HNO3 by microwave digestion before determination. The eight metal elements, potassium, nickel, calcium, magnesium, iron, manganese, copper and zinc, in Cichorium glandulosum Boiss et Huet were determined by FAAS. The working conditions, accuracy and precision of the method were studied. The linear correlations of standard curves are good (r = 0.999 1-0.999 9). The recovery (n = 6) is 92.25%-110.5%, and the RSD (n = 6) is 0.7%-3.88%. The results showed that there were comparatively rich metal elements, among which are comparatively high calcium (65.84 mg x g(-1)), iron (24.38 mg x g(-1)), magnesium (278.17 mg x g(-1)) and potassium (18.50 mg x g(-1)), in Cichorium glandulosum Boiss et Huet, and the contents of other elements are nickel of 0.004 38 mg x g(-1), manganese of 0.52 mg x g(-1), copper of 0.016 5 mg x g(-1) and zinc of 0.18 mg x g(-1). This provided useful data for discussing the relationship between the content of the metal elements in Cichorium glandulosum Boiss et Huet and its clinical application in cardiovascular and osteoporosis disease.

Bone-strengthening supplement (BSP) promotes bone and cartilage repair, for the treatment of Osteonecrosis of Femoral Head: an MRI-based study.

Currently, no effective drug treatment is available for bone and joint disease, a disorder of the bone and cartilage cells. Osteonecrosis of the femoral head (ONFH) is an example of bone and joint disease. It is progressive, with femoral head collapse resulting from the death of osteocytes and the bone marrow, leading to a poor quality of life and surgical interventions. However, the mechanism of this disease is still unknown, and the effects of current therapy are not satisfactory. In our previous study, we showed, using an ONFH rat model, that a new Chinese medicine, "bone-strengthening supplement" (BSP), enhances bone growth, promotes bone density, and restores blood circulation in the femoral head, and can significantly relieve pain, improve hip joint function, and reduce claudication. In the present study, we evaluated the curative effect of BSP in patients with ONFH using MRI with a double-blind randomized protocol. BSP significantly relieved pain unlike the control treatment; in addition, this treatment could improve MRI signal in ONFH patients. These results suggest that, overall; BSP can restore blood circulation and promote bone and cartilage growth during restoration of bone necrosis and the treatment of bone and joint disease.

Clomipramine causes osteoporosis by promoting osteoclastogenesis via E3 ligase Itch, which is prevented by Zoledronic acid.

Patients taking antidepressants, including Clomipramine (CLP), have an increased risk of osteoporotic fracture. However, the effects of CLP on bone metabolism are unknown. Here, we demonstrate that WT mice treated with CLP for 2 weeks had significantly reduced trabecular bone volume and cortical bone thickness, associated with increased osteoclast (OC) numbers, but had no change in osteoblast numbers or bone formation rate. Bone marrow cells from CLP-treated mice had normal OC precursor frequency, but formed significantly more OCs when they were cultured with RANKL and M-CSF. CLP promoted OC formation and bone resorption and expression of OC-associated genes. CLP-induced bone loss was prevented by Zoledronic acid. At the molecular level, CLP inhibited the activity of the ubiquitin E3 ligase Itch. CLP did not promote OC formation from bone marrow cells of Itch-/- mice in vitro nor induce bone loss in Itch-/- mice. Our findings indicate that CLP causes bone loss by enhancing Itch-mediated osteoclastogenesis, which was prevented by Zoledronic acid. Thus, anti-resorptive therapy could be used to prevent bone loss in patients taking antidepressants, such as CLP.

Bone-strengthening pill (BSP) promotes bone cell and chondrocyte repair, and the clinical and experimental study of BSP in the treatment of osteonecrosis of the femoral head.

About 1 in 3 people suffer from bone and joint disease, which is a disease of bone and cartilage cells. Osteonecrosis of the femoral head (ONFH) is a typical example of bone and joint disease involving bone cell necrosis. Osteonecrosis of the femoral head leads to the occurrence of premature osteoarthritis of the hip and collapse of the cartilage cells, and there is currently no effective drug treatment available. In order to study the effects of "bone-strengthening pill" (BSP) on the repair of bone and cartilage cells, we investigated the potential effects of the herbal mixture BSP in an animal model of avascular necrosis of the femoral head and in patients. Results showed that 90% of rats injected with prednisone developed ONFH, whereas BSP administration prevented ONFH development in 70% of prednisone-injected rats. We evaluated the constituents of BSP by HPLC fingerprinting. We also evaluated the clinical efficacy of BSP in a double-blind, randomized, controlled trial of 300 patients with ONFH. The response rate was found to be higher in the treatment group than in the control group, with a response rate of 82% in the treatment group. Treatment with BSP also significantly reduced pain, improved hip function, reduced lameness, and improved pathology by X-ray and MRI analysis, compared with patients who did not receive BSP. These results suggest that BSP treatment inhibits and reverses necrosis of the femoral head bone cells and cartilage cells to repair the femoral head, promote the repair of bone and cartilage diseases.

[Influence of aqueous extract of Aralia echinocaulis Hand.-Mazz on the expression of fracture healing-related factor receptors].

OBJECTIVE: To investigate the influence of aqueous extract of Aralia echinocaulis Hand.-Mazz on the expression of fracture healing-ralated factor receptors. METHODS: Single factor model was set up in SD rat. Selecting 14 and 28 days in the experiment. Immunohistochemistry was employed to determine the expression of Fibroblast growth factor receptor 2 (FGFR2), Fms-like tyrosine kinase (Flt-1) and Fetal licer kinase (Flk-1) at 14 and 28 days after model establishing. RESULTS: The expression of Flt-1 and Flk-1 at 14 days (the latter was more remarkable) were obviously promoted in High dose group of aqueous extract of Aralia echinocaulis Hand.-Mazz, and higher than that in normal group and model group. The expression of FGFR2 in the high dose group of Aralia echinocaulis Hand -Mazz was also promoted visibility, close to that in the compare group (traditional Chinese medicine), but higher than than in the model group. There was no significant difference among them. At 28 days, the expression of FGFR2, Flt-1 and Flk-1 in all groups decreased except normal group, and got higher expression in model groups than each control groups. CONCLUSION: Aqueous extract of Aralia echinocaulis Hand.-Mazz can promote angiogenesis in fracture healing, improve the activity and aggregation of fibroblasts, osteoblasts and chondrocytes. It also helps to quicken ossification in the cartilage and promote fracture healing.

[Effects of Astaxanthin on the damage of osteoblast induced by H2O2].

OBJECTIVE: To investigate the effect of Astaxanthin on enhancing the function of anti-oxidative damage in osteoblast. METHODS: MC3T3-E1 osteoblasts were randomly divided into five groups, including control group, model group, Astaxanthin group [low-dose (1 x 10(-7) mol/L), middle-dose (1 x 10(-6) mol/L), high-dose (1 x 10(-5) mol/L)], in which the activity of cells, activity of superoxide dismutase (SOD), the content of reactive oxygen species (ROS), lipid oxygen (LPO) and membrane fluidity were tested and compared. RESULTS: Compared with Astaxanthin groups, the activity of cells, SOD activity and membrane fluidity in the model group were significantly decreased (P < 0.01). However, the contents of ROS and LPO were significantly raised (P < 0.01). CONCLUSION: H2O2 can cause oxidative damage of MC3T3-E1 osteoblasts, but Astaxanthin can prevent or decrease its influence.

[Influence of canthaxanthin on D-galactose induced osseous changes of rat].

OBJECTIVE: To study the influence of canthaxanthin on D-galactose induced osseous changes of rat. METHODS: Forty-five six-week-old Wistar male rats were randomly divided into model group, canthaxanthin group and young control group. In addition, 15 sixteen-month-old Wistar male rats were used as old control group. Model group and canthaxanthin group were given injections of D-galactose for 5 months (20 mg/kg/once per-day) to cause aging of rat. Then routine osseous parameters were tested and compared among the 4 groups. RESULTS: Compared with young control group, the BMD, parameters of structural mechanics and biomechanics, bone calcium, manganese, magnesium and the content of hydroxyproline in the model group decreased significantly (P < 0.01), however, the content of bone phosphorus, the activity of bone and serum ALP increased significantly (P < 0.01). Those changes of the model group were the same as the old control group,but the changes in the canthaxanthin group significantly differed with the model group (P < 0.01). CONCLUSION: The high does of D-galactose intake can cause aging and osteoporosis at the same time in rat, but canthaxanthin can prevent and inhibit D-galactose induced osseous changes.