Bisphosphonate-enoxacin inhibit osteoclast formation and function by abrogating RANKL-induced JNK signalling pathways during osteoporosis treatment.

Osteoporosis is an age-related disease characterized by low mineral density, compromised bone strength and increased risk of fragility fracture. Most agents for treating osteoporosis focus primarily on anti-resorption by inhibiting osteoclast activity. Bisphosphonate (BP) is a potent anti-resorptive agent that has been used clinically for decades and is proven to be effective. However, BP has a variety of side effects and is far from being an ideal anti-osteoporosis agent. BP selectively binds to calcium crystals, which are subsequently taken up or released by osteoclasts. Based on the action of BP, we previously demonstrated the inhibitory effect of a novel bone-targeting BP derivative, bisphosphonate-enoxacin (BE). In the current study, we used bone marrow-derived osteoclast cultures to further assess the inhibitory effect of BE on osteoclastogenesis and employed reverse transcription PCR and real-time PCR to examine expression of osteoclast-specific genes. Additionally, we used bone resorption and F-actin immunofluorescence assays to evaluate the effect of BE on osteoclast function and investigated the potential mechanisms affecting osteoclast differentiation and function in vitro. Furthermore, an ovariectomized (OVX) rat model was established to evaluate the therapeutic effects of BE on preventing bone loss. Results showed that BE exerted potent inhibitory effects on osteoclast formation and bone resorption by specifically abrogating RANKL-induced JNK signalling, and that it preserved OVX rat bone mass in vivo without any notable side effects. Collectively, these results indicated that the BP derivative BE may have significant potential as a treatment for osteoporosis and other osteolytic diseases.

Analyzing the molecular mechanism of the tissue specificity of gastrointestinal stromal tumors by using bioinformatics approaches.

PURPOSE: Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. About 70% of GIST occur in the stomach, 20% in the small intestine and less than 10% in the esophagus. This study aimed to explore the difference of gene expression profile of GIST between different tumor sites. METHODS: Microarray data GSE8167 (accession number of the microarray data) were available from Gene Expression Omnibus (GEO) which included 23 gastric and 9 small intestine untreated GIST samples, and then the differentially expressed genes (DEGs) between these samples were identified using t-test. Furthermore, pathway enrichment analysis was performed to these DEGs and one protein-protein interaction network was constructed by STRING. Additionally, BioNet in R was used to establish a sub-network with false discovery rate < 0.001, and genes in this sub-network were further subjected to gene ontology (GO) and pathway analyses. RESULTS: A total 730 genes were differentially expressed between gastric samples and small intestine samples, indicating the tissue specifity of GIST. Pathway analysis suggested these DEGs disturbed ECM-receptor interaction, gap junction and colorectal cancer. Moreover, some nodes (such as PLAT, VEGFC, PGF and CHD7) in the sub-network were significantly enriched in blood vessel development (p=4.58E-06), appendage development (p=9.54E-06) and skeletal system development (p=2.40E-04), respectively. Finally, several DEGs in the sub-network, including VEGFC and PGF, mainly affected pathways in cancer, focal adhesion, bladder cancer and cytokine-cytokine receptor interaction. CONCLUSIONS: Our results suggest that molecular mechanisms of GIST originating in different site were different. Our findings are helpful for physicians and researchers to study the tissue specificity of GIST.

Expression and significance of DDX43 in lung adenocarcinoma.

This paper aims to determine the expression and clinical significance of DDX43 in lung adenocarcinoma. Expression of DDX43 gene and protein of lung adenocarcinoma tissue and para-carcinoma tissues was observed in 27 cases by RT-PCR and immunohistochemistry. These patients were diagnosed as lung adenocarcinoma in the Huaihe Hospital of Henan University from February 2015 to December 2015. The relative ratio of DDX43 mRNA expression in lung adenocarcinoma and para-carcinoma tissues was 0.87±0.62 versus 0.21±0.77 and the difference between the two groups was statistically significant (P<0.01). The expression of DDX43 in normal lung tissues and lung adenocarcinoma tissues was different. The positive rate of DDX43 expression in lung adenocarcinoma tissues was significantly higher than that in normal lung tissues, and the difference was statistically significant (P<0.05). The analysis of clinical pathological characteristics showed that the increase of protein expression was related to the stage and metastasis of lung adenocarcinoma. DDX43 is highly expressed in lung adenocarcinoma, and the expression level is related to the stage and metastasis of lung adenocarcinoma, suggesting that DDX43 is closely related to the occurrence and development of lung adenocarcinoma, and may be a molecular marker for early diagnosis of lung adenocarcinoma.

Incorporation of cerium oxide into zirconia toughened alumina ceramic promotes osteogenic differentiation and osseointegration.

Due to its high wear resistance and good biocompatibility, zirconia toughened alumina (ZTA) is an ideal material used as load-bearing implant. However, ZTA needs to be modified to overcome its bio-inert and thus improve osseointegration. Cerium oxide, which has been proved to be a bone-friendly ceramic, might be a desired material to enhance the bioactivity of ZTA. In this study, ZTA and cerium oxide doped ZTA (ZTAC) were prepared via sintering method. The in vitro study showed that the addition of cerium oxide promoted MC3T3-E1 cell adhesion and spreading through upregulating ITG α5 and ITG ß1. In addition, the incorporation of cerium oxide enhanced cell proliferation, ALP activity, and ECM mineralization capacity. Moreover, the incorporation of cerium oxide promoted the expressions of osteogenesis related genes, such as ALP, Col-I, and OCN. The in vivo implantation test via a SD rat model showed that the incorporation of cerium oxide promoted new bone formation and bone-implant integration. In summary, this study provided a new strategy to fabricate bioactive ZTA implant for potential application in orthopedics field.

Celastrol Attenuates RANKL-Induced Osteoclastogenesis in vitro and Reduces Titanium Particle-Induced Osteolysis and Ovariectomy-Induced Bone Loss in vivo.

Excessive bone resorption by osteoclasts contributes significantly to osteoclast-related diseases such as periprosthetic osteolysis and osteoporosis. Osteolysis in a titanium particle-induced calvarial model and bone loss in an ovariectomized mice model occurred similarly to those in humans; thus, these models can be used to evaluate potential therapies for aseptic prosthetic loosening and osteoporosis. Celastrol, which is extracted from the seeds of the genus Tripterygium, has been thoroughly investigated for its anti-inflammatory and anti-cancer pharmacological effects. However, the mechanisms involving bone metabolism by which celastrol inhibits osteoclastogenesis are not yet fully understood. We demonstrated that celastrol inhibited the receptor activator of nuclear factor κB ligand-induced osteoclastogenesis and the bone resorptive function of osteoclasts in vitro by inhibiting the activation of transforming growth factor ß-activated kinase 1-mediated NF-κB and mitogen-activated protein kinase signaling pathways and downregulating osteoclastogenesis marker-related genes. Furthermore, celastrol was also shown to be beneficial in both the titanium particle-induced osteolysis calvarial and the murine ovariectomy-induced bone loss. Collectively, our results suggested that celastrol is promising for the prevention of aseptic prosthetic loosening and osteoporosis in the treatment of osteolytic diseases induced by disrupted osteoclast formation and function.

Glaucocalyxin A suppresses osteoclastogenesis induced by RANKL and osteoporosis induced by ovariectomy by inhibiting the NF-κB and Akt pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Glaucocalyxin A (GLA), the most abundant active component of the aboveground sections of Rabdosia japonica (Burm. f.) Hara var. glaucocalyx (Maxim.) Hara, possesses various pharmacological activities, such as antioxidant, antithrombosis, anticoagulation, antibacterial, antitumor, anti-inflammatory activities. According to previous studies, inflammation is closely associated with osteoclast differentiation and activity. Although GLA has demonstrated effective anti-inflammatory properties, its effects on osteoclast differentiation remain unclear. AIM OF THE STUDY: To examine the possible inhibitory effects of GLA and its molecular mechanisms in osteogenesis induced by RANKL as well as ovariectomy (OVX)-induced osteoporosis (OP) in mice. MATERIALS AND METHODS: Tartrate-resistant acid phosphatase (TRAP) staining, F-actin staining, and a bone resorption pit assay were applied for identifying the effects of GLA on the differentiation of osteoclasts and the function of bone resorption. The mRNA expression of the genes related to osteoclast differentiation was measured by quantitative PCR. Protein expression of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), c-fos and phosphorylation of inhibitor of nuclear factor kappa B (IκBα), protein kinase B (AKT), c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 in RANKL-induced osteoclasts was determined using western blotting. The effect of GLA on OP was studied using a mouse model of OVX. RESULTS: At nontoxic concentrations ≤0.5 µM in vitro, GLA suppressed the formation of osteoclasts induced by RANKL with the decreased number and area size of TRAP-positive multinuclear osteoclasts, and the resorption of bone function by reducing F-actin ring number and bone resorption pit areas. It also reduced the expression of the genes specific for osteoclasts, which included genes encoding NFATc1, cathepsin K, c-fos, TRAP, vacuolar-type ATPase d2, and dendritic cell-specific transmembrane protein. Moreover, GLA repressed NF-κB and Akt pathway activation induced by RANKL. Micro-CT analysis of femur samples indicated decreased bone loss and greater trabecular bone density after GLA treatment, which showed that GLA played a protective role by inhibiting bone loss in OVX-induced OP mice in vivo. CONCLUSIONS: Our study is the first to show that GLA has significant therapeutic potential in OP, which is the disease of osteoclast increase caused by estrogen deficiency.

Hospital length of stay following first-time elective open posterior lumbar fusion in elderly patients: a retrospective analysis of the associated clinical factors.

To identify independent factors associated with prolonged hospital length of stay (LOS) in elderly patients undergoing first-time elective open posterior lumbar fusion surgery.We retrospectively analyzed the data of 303 elderly patients (age range: 60-86 years) who underwent first-time elective open lumbar posterior fusion surgery at our center from December 2012 to December 2017. Preoperative and perioperative variables were extracted and analyzed for all patients, and multivariate stepwise regression analysis was used to determine the variables affecting the LOS and important predictors of LOS prolongation (P < .001).The mean age of the patients was 67.0 ±â€Š5.5 years, and the mean LOS was 18.5 ±â€Š11.8 days, ranging from 7 to 103 days. Of the total, 166 patients (54.8%) were men and 83 patients (27.4%) had extended LOS. Multiple linear regression analysis determined that age (P < .001), preoperative waiting time ≥7 days (P < .001), pulmonary comorbidities (P = .010), and diabetes (P = .010) were preoperative factors associated with LOS prolongation. Major complications (P = .002), infectious complications (P = .001), multiple surgeries (P < .001), and surgical bleeding (P = .018) were perioperative factors associated with LOS prolongation. Age (P < .001), preoperative waiting time ≥7 days (P < .001), infectious complications (P < .001), and multiple surgeries (P < .001) were important predictors of LOS prolongation.Extended LOS after first-time elective open posterior lumbar fusion surgery in elderly patients is associated with factors including age, preoperative waiting time, infectious complications, and multiple surgeries. Surgeons should recognize and note these relevant factors while taking appropriate precautions to optimize the modifiable factors, thereby reducing the LOS as well as hospitalization costs.

Simultaneous ABO-incompatible living-donor liver transplantation and splenectomy without plasma exchange in China: Two case reports.

ABO-incompatible (ABO-i) living-donor liver transplantation (LDLT) is performed if an ABO-compatible graft cannot be obtained. However, a perfect desensitization protocol has not been established worldwide, especially for simultaneous ABO-i LDLT and splenectomy. We herein report two cases of ABO-i LDLT. To the best of our knowledge, this is the first case report of ABO-i LDLT in an adult patient in China. Splenectomy and T-cell-targeted immunosuppression (basiliximab) was used to overcome the blood group barrier in these recipients. The patients had good graft function without signs of antibody-mediated rejection throughout the 12-month follow-up. Thus, ABO-i LDLT with splenectomy is undoubtedly life-saving when an ABO-compatible graft cannot be obtained for patients in critical condition.