Targeting excessive osteoclast differentiation and activity is considered a valid therapeutic approach for osteoporosis. Zoledronic acid (Zol) plays a pivotal role in regulating bone mineral density. However, the exact molecular mechanisms responsible for the inhibitory effects of Zol on receptor activator of nuclear factor (nF)-κB ligand (ranKl)-induced osteoclast formation are not entirely clear. The present study aimed to investigate the role of Zol in osteoclast differentiation and function, and to determine whether nF-κB and mitogen-activated protein kinase, and their downstream signalling pathways, are involved in this process. raW264.7 cells were cultured with ranKl for differentiation into osteoclasts, in either the presence or absence of Zol. osteoclast formation was observed by tartrate-resistant acid phosphatase staining and bone resorption pit assays using dentine slices. The expression of osteoclast-specific molecules was analysed using reverse transcription-quantitative polymerase chain reaction and western blotting assays to deduce the molecular mechanisms underlying the role of Zol in osteoclastogenesis. The results showed that ZOL significantly attenuated osteoclastogenesis and bone resorptive capacity in vitro. Zol also suppressed the activation of nF-κB and the phosphorylation of c-Jun n-terminal kinase. Furthermore, it inhibited the expression of the downstream factors c-Jun, c-Fos and nuclear factor of activated T cells c1, thereby decreasing the expression of dendritic cell-specific transmembrane protein and other osteoclast-specific markers. in conclusion, Zol may have therapeutic potential for osteoporosis.

Bone mass loss (osteoporosis) seen in postmenopausal women is an adverse factor for implant denture. Using an ovariectomized rat model, we studied the mechanism of estrogen-deficiency-caused bone loss and the therapeutic effect of Zoledronic acid. We observed that ovariectomized-caused resorption of bone tissue in the mandible was evident at four weeks and had not fully recovered by 12 weeks post-ovariectomized compared with the sham-operated controls. Further evaluation with a TUNEL assay showed ovariectomized enhanced apoptosis of osteoblasts but inhibited apoptosis of osteoclasts in the mandible. Zoledronic acid given subcutaneously as a single low dose was shown to counteract both of these ovariectomized effects. Immunohistochemical staining showed that ovariectomized induced the protein levels of RANKL and the 65-kD subunit of the NF-κB complex mainly in osteoclasts, as confirmed by staining for TRAP, a marker for osteoclasts, whereas zoledronic acid inhibited these inductions. Western blotting showed that the levels of RANKL, p65, as well as the phosphorylated form of p65, and IκB-α were all higher in the ovariectomized group than in the sham and ovariectomized + zoledronic acid groups at both the 4th- and 12th-week time points in the mandible. These data collectively suggest that ovariectomized causes bone mass loss by enhancing apoptosis of osteoblasts and inhibiting apoptosis of osteoclasts. In osteoclasts, these cellular effects may be achieved by activating RANKL-NF-κB signalling. Moreover, zoledronic acid elicits its therapeutic effects in the mandible by counteracting these cellular and molecular consequences of ovariectomized.

BackgroundClostridium difficile infection (CDI) is the leading cause of nosocomial diarrhea. Co-colonization of key bacterial taxa may prevent the transition from asymptomatic C. difficile colonization to CDI. However, little is known about the composition of key bacterial taxa in asymptomatic patients.MethodsIn the present study, the culture method was used to examine the composition of stool microbiota in two asymptomatic patients from Guizhou, China.ResultsA total of 111 strains were isolated and phylogenetic relationships were determined by 16S ribosomal gene sequencing and Molecular Evolutionary Genetics Analysis version 7. The results demonstrated that Escherichia (33.3%, 37/111), Clostridium (24.3%, 27/111) and Enterococcus (11.7%, 13/111) exhibited a high ratio in asymptomatic patients. These isolates derived from two phyla: Firmicutes (51.3%, 57/111) and Proteobacteria (44.1%, 49/111). In addition, co-colonization of human pathogens Fusobacterium nucleatum, Ralstonia pickettii, Klebsiella pneumoniae, Klebsiella quasipneumoniae and Clostridium tertium with C. difficile was identified. To the best of our knowledge, these pathogens have not been co-isolated with C. difficile previously.ConclusionsIn summary, the present study identified the composition of fecal microbiota in two asymptomatic patients in Guizhou, China. These results suggested that co-infection with human pathogens may be ubiquitous during CDI progression.