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.

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 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.

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.

[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.