ML216-Induced BLM Helicase Inhibition Sensitizes PCa Cells to the DNA-Crosslinking Agent Cisplatin.

Using standard DNA-damaging medicines with DNA repair inhibitors is a promising anticancer tool to achieve better therapeutic responses and reduce therapy-related side effects. Cell viability assay, neutral comet assay, western blotting (WB), and cell cycle and apoptosis analysis were used to determine the synergistic effect and mechanism of ML216, a Bloom syndrome protein (BLM) helicase inhibitor, and cisplatin (CDDP), a DNA-crosslinking agent, in PCa cells. Based on the online database research, our findings revealed that BLM was substantially expressed in PCa, which is associated with a bad prognosis for PCa patients. The combination of ML216 and CDDP improved the antiproliferative properties of three PCa cell lines. As indicated by the increased production of γH2AX and caspase-3 cleavage, ML216 significantly reduced the DNA damage-induced high expression of BLM, making PC3 more susceptible to apoptosis and DNA damage caused by CDDP. Furthermore, the combination of ML216 and CDDP increased p-Chk1 and p-Chk2 expression. The DNA damage may have triggered the ATR-Chk1 and ATM-Chk2 pathways simultaneously. Our results demonstrated that ML216 and CDDP combination therapy exhibited synergistic effects, and combination chemotherapy could be a novel anticancer tactic.

Enzymatic Reaction Generates Biomimic Nanominerals with Superior Bioactivity.

In vivo mineralization is a multistep process involving mineral-protein complexes and various metastable compounds in vertebrates. In this complex process, the minerals produced in the mitochondrial matrix play a critical role in initiating extracellular mineralization. However, the functional mechanisms of the mitochondrial minerals are still a mystery. Herein, an in vitro enzymatic reaction strategy is reported for the generation of biomimic amorphous calcium phosphate (EACP) nanominerals by an alkaline phosphatase (ALP)-catalyzed hydrolysis of adenosine triphosphate (ATP) in a weakly alkalescent aqueous condition (pH 8.0-8.5), which is partially similar to the mitochondrial environment. Significantly, the EACP nanomineral obviously promotes autophagy and osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by activating an AMPK related pathway, and displays a high performance in promoting bone regeneration. These results provide in vitro evidence for the effect of ATP on the formation and stabilization of the mineral in the mineralization process, demonstrating a potential strategy for the preparation of the biomimic mineral for treating bone related diseases.

Evaluation of Quality of Life Using EORTC QLQ-BM22 in Patients with Bone Metastases after Treatment with Magnetic Resonance Guided Focused Ultrasound.

OBJECTIVE: To reveal the alterations in quality of life (QOL) in bone metastases patients after magnetic resonance guided focused ultrasound (MRgFUS). METHODS: This retrospective study enrolled 26 patients diagnosed with bone metastases. Patients had various primary malignant tumors and tumor lesions in different locations. All patients received MRgFUS for bone metastasis. Each focal spot sonication pulse that was applied to create energy deposition lasted 20 s and was performed at a frequency of 1.05 MHz. The visual analog scale (VAS) was used to measure pain level and the EORTC QLQ-BM22 was applied to evaluate QOL for 12 months. The lower the QLQ-BM22 score, the better the QOL of patients. RESULTS: The painful site subscale of the EORTC QLQ-BM22 was observed without significant change. Significant reductions in the functional subscales were observed after therapy compared with the baseline. The functional interference was reduced significantly during the first 12 months. From the 2-month time point onwards, the pain characteristics subscale also decreased significantly. VAS scores had decreased by 40.8% 1 month after the operation and had decreased 10.9% compared with VAS scores preoperation. Scores for pain characteristics decreased by 28.8% after the operation and the scores were still down by 10.8% 1 year after the treatment. VAS scores indicated a significant reduction in pain over the course of the research until the 12-month time point follow-up compared with the baseline. CONCLUSION: MRgFUS therapy improved the QOL of patients with bone metastasis by relieving bone pain.

Calcium phosphate-phosphorylated adenosine hybrid microspheres for anti-osteosarcoma drug delivery and osteogenic differentiation.

Biocompatibility, biodegradability and bioactivity are significantly important in practical applications of various biomaterials for bone tissue engineering. Herein, we develop a functional inorganic-organic hybrid system of calcium phosphate-phosphorylated adenosine (CPPA). Both calcium phosphate and phosphorylated adenosine molecules in CPPA are fundamental components in mammalians and play important roles in biological metabolism. In this work, we report our three leading research qualities: (1) CPPA hybrid microspheres with hollow and porous structure are synthesized by a facile one-step microwave-assisted solvothermal method; (2) CPPA hybrid microspheres show high doxorubicin loading capacity and pH-responsive drug release properties, and demonstrate positive therapeutic effects on six osteosarcoma cell lines in vitro and a mouse model of 143B osteosarcoma subcutaneous tumor in vivo; (3) CPPA hybrid microspheres are favorable to promote osteogenic differentiation of human bone mesenchymal stem cells (hBMSCs) by activating the AMPK pathway, with satisfactory evidences from cellular alkaline phosphatase staining, alizarin red staining, real time PCR and western analysis. The as-prepared CPPA hybrid microspheres are promising in anti-osteosarcoma and bone regeneration, which simultaneously display excellent properties on drug delivery and osteogenic differentiation of hBMSCs.

Surgical Treatment of Sacral Neurogenic Tumor: A 10-year Experience with 64 Cases.

OBJECTIVE: To explore the outcomes of surgical treatment of sacral neurogenic tumors METHODS: Between 1 January 2003 and 31 December 2012, data on 64 patients with sacral neurogenic tumors treated with surgery were retrospectively analyzed. The mean age of the 64 cases (35 males and 29 females) was 37.2 years (range, 21-69 years); 38 had neurilemmomas and 26 neurofibromas. Thirty-four of the tumors involved S 1 and S 2 , 11 S 3 or lower, and 19 were single presacral soft tissue masses. Tumors were removed via anterior, posterior or combined anteroposterior approaches. Patients with unstable sacroiliac joints underwent iliolumbar fixation. RESULTS: Depending on the extent of tumor involvement, one of three surgical approaches was used: a single anterior approach (19 patients), single posterior approach (25 patients), or a combined anteroposterior approach (20 patients). The mean operation time was 3 h (range, 2-6 h) and the mean blood loss 878 mL (range, 400-3120 mL). The mean duration of follow-up was 58.2 months (range, 24-93 months). These surgeries had the following complications. Three patients had massive intraoperative hemorrhage and posterior back pain and discomfort postoperatively. One patient had intraoperative ureteral injuries requiring intraoperative ureteral catheterization. In two patients, the tumor involved the S 1 nerve roots bilaterally, necessitating their removal, which resulted in obvious lower limb motion and sphincteric dysfunction. In 13 patients with unilateral tumor involvement of the nerve roots of S 1 and lower spinal levels, only the contralateral nerve roots of the S1 and lower levels were preserved; eight of these patients had impaired bladder and bowel function. Posterior incisions failed to heal in 10 patients, secondary wound healing occurred in nine of them and one required a gluteus maximus myocutaneous flap. Three patients developed postoperative cerebrospinal fluid leaks that were and alleviated by waist belt compression bandaging and placing them in the Trendelenburg position. Eight patients developed tumor recurrences postoperatively; pathological examination of the tissue excised in the second surgeries revealed malignant changes in the three patients with neurilemmomas. There were no intraoperative deaths. Rod fractures occurred in three of the 18 patients requiring iliolumbar reconstruction. CONCLUSIONS: The clinical characteristics of sacral neurogenic tumors make them easy to diagnose. The approach to resection should be determined by the location and size of the tumor. Patients with huge tumors may lose considerable blood intraoperatively and a have higher risk rate of postoperative complications.

AMD3100 reduces CXCR4-mediated survival and metastasis of osteosarcoma by inhibiting JNK and Akt, but not p38 or Erk1/2, pathways in in vitro and mouse experiments.

Osteosarcoma (OS) has an unfavorable prognosis and tends to metastasize to lung tissue. Although the CXCL12-CXCR4 axis appears to affect progression and metastasis in numerous tumors, its mechanism and downstream pathways in OS remain unclear. We used western blotting and flow cytometry to detect CXCR4 and CXCR7 expression in two OS cell lines (LM8 and Dunn). An MTT assay was used to evaluate the effects of CXCL12 and AMD3100, a specific CXCR4 antagonist, on cell viability. Flow cytometry was utilized to analyze changes in apoptosis induced by serum deprivation following treatment with CXCL12 and AMD3100. A Transwell assay was used to assess cell migration in response to CXCL12 and AMD3100. Western blotting was performed to identify the phosphorylation of signaling molecules (JNK, c-Jun, Akt, p38 and Erk1/2) and expression of caspase-3 and -8, and PARP. Mouse models were employed to evaluate AMD3100 inhibition of primary OS growth and lung metastasis in vivo. CXCR4 expression was detected in LM8 but not Dunn cells, and neither cell line expressed CXCR7. The addition of CXCL12 induced the survival and migration of serum-starved CXCR4+ LM8 cells activating JNK and Akt pathways, which were abrogated by adding AMD3100. However, similar results were not observed in CXCR4- Dunn cells. CXCL12 protected LM8, but not Dunn cells, from apoptosis induced by serum deprivation by suppressing PARP cleavage, which was partly reversed by AMD3100. In a mouse model, AMD3100 reduced primary tumor growth and lung metastasis compared with the controls. Thus, the CXCL12-CXCR4 axis regulated OS survival and metastasis through the JNK and Akt pathways, and blocking them with AMD3100 was found to be a potential OS treatment.

Prognostic significance of serum lactate dehydrogenase level in osteosarcoma: a meta-analysis.

BACKGROUND: A number of studies have investigated the role of serum lactate dehydrogenase (LDH) level in patients with osteosarcoma but have yielded inconsistent and inconclusive results. Thus, we conducted a meta-analysis to assess its prognostic value more precisely. METHODS: Systematic computerized searches of PubMed, Embase and Web of Science databases were performed. The pooled hazard ratio (HR) with 95 % confidence intervals (95 % CI) of overall survival was used to assess the prognostic role of serum LDH level. RESULTS: Ten studies published between 1997 and 2013 with a total of 943 osteosarcoma patients were included. Overall, the pooled HR for all ten eligible studies evaluating high LDH level on overall survival was 1.92(95 % CI 1.53-2.40). Sensitivity analysis suggested that the pooled HR was stable and omitting a single study did not change the significance of the pooled HR. Funnel plots and Egger's tests revealed there was some possibility of publication bias risk in the meta-analysis. CONCLUSION: This meta-analysis shows that high serum LDH level is obviously associated with lower overall survival rate in patients with osteosarcoma, and it is an effective biomarker of prognosis.

Establishing an osteosarcoma associated protein-protein interaction network to explore the pathogenesis of osteosarcoma.

BACKGROUND: The aim of this study was to establish an osteosarcoma (OS) associated protein-protein interaction network and explore the pathogenesis of osteosarcoma. METHODS: The gene expression profile GSE9508 was downloaded from the Gene Expression Omnibus database, including five samples of non-malignant bone (the control), seven samples for non-metastatic patients (six of which were analyzed in duplicate), and 11 samples for metastatic patients (10 of which were analyzed in duplicate). Differentially expressed genes (DEGs) between osteosarcoma and control samples were identified by packages in R with the threshold of |logFC (fold change)| > 1 and false discovery rate < 0.05. Osprey software was used to construct the interaction network of DEGs, and genes at protein-protein interaction (PPI) nodes with high degrees were identified. The Database for Annotation, Visualization and Integrated Discovery and WebGestalt software were then used to perform functional annotation and pathway enrichment analyses for PPI networks, in which P < 0.05 was considered statistically significant. RESULTS: Compared to the control samples, the expressions of 42 and 341 genes were altered in non-metastatic OS and metastatic OS samples, respectively. A total of 15 significantly enriched functions were obtained with Gene Ontology analysis (P < 0.05). The DEGs were classified and significantly enriched in three pathways, including the tricarboxylic acid cycle, lysosome and axon guidance. Genes such as HRAS, IDH3A, ATP6ap1, ATP6V0D2, SEMA3F and SEMA3A were involved in the enriched pathways. CONCLUSIONS: The hub genes from metastatic OS samples are not only bio-markers of OS, but also help to improve therapies for OS.

The role of the CXCL12-CXCR4/CXCR7 axis in the progression and metastasis of bone sarcomas (Review).

Bone sarcomas, which comprise less than 1% of all human malignancies, are a group of relatively rare mesenchymal-derived tumors. They are mainly composed of osteosarcoma, chondrosarcoma and Ewing's sarcoma. In spite of advances in adjuvant chemotherapy and wide surgical resection, prognosis remains poor due to the high propensity for lung metastasis, which is the leading cause of mortality in patients with bone sarcomas. Chemokines are a superfamily of small pro-inflammatory chemoattractant cytokines which can bind to specific G protein-coupled seven-span transmembrane receptors. Chemokine 12 (CXCL12), also designated as stromal cell-derived factor-1 (SDF-1), is able to bind to its cognate receptors, chemokine receptor 4 (CXCR4) and chemokine receptor 7 (CXCR7), with high affinity. The binding of CXCL12 to CXCR4/CXCR7 stimulates the activation of several downstream signaling pathways that regulate tumor progression and metastasis. In this review, the structure and function of CXCL12 and its receptors, CXCR4 and CXCR7, as well as many factors affecting their expression are discussed. Phosphoinositide 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) pathways are the two most important downstream pathways regulated by the CXCL12-CXCR4/CXCR7 interaction. CXCR4 expression in bone sarcomas, including tumor cells and samples and the correlation between CXCR4/CXCR7 expression and the survival of patients with bone sarcomas are also discussed. In addition, we review the involvement of the CXCL12­CXCR4/CXCR7 axis in the growth and metastasis of bone sarcomas and the targeting of this axis in preclinical studies.

Determination of the apoptotic index in osteosarcoma tissue and its relationship with patients prognosis.

BACKGROUND: Nowadays it remains a controversial issue whether a correlation exists between the apoptosis rate of tumor tissue and the prognosis of the patients. We aimed to explore the prognostic significance of apoptosis index of human osteosarcoma tissue. METHODS: The technique of terminal DNA breakpoints in situ 3 - hydroxy end labeling (TUNEL) was used to detect and analysis apoptosis index in 56 osteosarcoma specimens. The relationships between apoptosis index of tumor tissue and long term survival of patients as well as pathologic classification, tumor clinical stages, tumor size and level of serum alkaline phosphatase were analyzed. RESULTS: Our studies showed the cases with high apoptosis index had significantly longer survival time. Apoptosis index in osteosarcoma tissue was correlated with tumor size and level of serum alkaline phosphatase but not with pathologic classifications and clinical stages of tumor. CONCLUSION: Our results demonstrated that apoptosis index of osteosarcoma tissue combined with serum alkaline phosphatase could used as valid indicators to predicate the malignant level and prognosis of osteosarcoma cases, which would contribute to enhance efficacy of clinical treatments for osteosarcoma.

Benzochloroporphyrin derivative induced cytotoxicity and inhibition of tumor recurrence during photodynamic therapy for osteosarcoma.

Photodynamic therapy (PDT) is a promising cancer treatment modality that uses dye-sensitized photooxidation of biologic matter in target tissue. This study explored effects of the photosensitizer BCPD-17 during PDT for osteosarcoma. LM-8 osteosarcoma cells were treated with BCPD-17 and cell viability after laser irradiation was assessed in vitro with the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. The effects of BCPD-17 during PDT recurrence were then examined on tumor-bearing mice in vivo. BCPD-17 had dose- dependent cytotoxic effects on LM-8 osteosarcoma cells after laser irradiation which also had energy-dependent effects on the cells. The rate of local recurrence was reduced when marginal resection of mice tumors was followed by BCPD-17-mediated PDT. Our results indicated BCPD-17-mediated PDT in combination with marginal resection of tumors is a potentially new effective treatment for osteosarcoma.

Telangiectatic osteosarcoma: a review of literature.

Telangiectatic osteosarcoma is a rare variant of osteosarcoma and hence its occurrence, presentation, and prognosis are poorly understood. With advancements in technology and available treatment options, the scenario of its diagnosis, management, and outcome has changed. Chemotherapy with surgery was challenged previously, but has now been proved to be beneficial. We reviewed the available literature and compared results to define the characteristics of the disease, its presentation, radiographic and pathologic features, optimal treatment, and prognosis.

The effects of inhibiting hedgehog signaling pathways by using specific antagonist cyclopamine on the chondrogenic differentiation of mesenchymal stem cells.

This study aimed to investigate the effects of cyclopamine, a specific inhibitor of Hedgehog signaling pathways, on the chondrogenic differentiation of mesenchymal stem cells (MSCs). During culture, the experimental groups were treated with cyclopamine and their cell proliferation status was assessed using the MTT test. The extra-bone cellular matrix (ECM) and Collagen II (Col II) was detected by toluidine blue staining and immunohistochemistry of cells. The concentrations of Col II and aggrecan in the culture solution and cytosol were detected using ELISA on the 7th, 14th, and 21st days of cyclopamine induction. Gene and protein expression of Col II and aggrecan were analyzed on the 14th day of cyclopamine induction using real-time PCR and western blot analyses. No significant differences in proliferation of mesenchymal stem cells were found between the control group and the group treated with cyclopamine. Compared to the blank control group, the ECM level was low and the protein and mRNA concentrations of Collagen II (Col II) and aggrecan in the culture solution and cytosol, respectively, were significantly reduced in the experimental group. The Smo acted as a key point in the regulations of Hedgehog signaling pathway on the chondrogenic differentiation of rabbit MSCs.

Treatments for acute and old distal radius fracture with lunate dislocation.

PURPOSE: Our study was undertaken to determine the correct treatment protocol for distal radius fracture with lunate anterior dislocation. METHODS: From 2000 to 2007, 58 patients (36 with acute injury and 22 with old injury) with distal radius fracture with lunate anterior dislocation were enrolled in the study. Among acute injury patients, 15 were treated through manipulative reduction and plaster fixation and 21 were treated through minimal invasive poking reduction followed by Kirschner wire and plaster fixation. Among old injury patients, 8 underwent operative reduction of lunate dislocation through the palmar approach and Kirschner wire and plaster fixation, whereas 14 patients underwent operative reduction and fixation through the dorsal approach combined with reparation of the dorsal radiolunate ligament. Lidstrom wrist function scores and the morbidity of lunate necrosis and osteoarthritis were documented and evaluated. RESULTS: Lidstrom wrist function scores revealed that the rate of excellent and good scores was higher in acute injury patients than in old injury patients (91.7 versus 54.5%, respectively; P = 0.018). The lunate necrosis rate was lower in acute injury patients than in old injury patients (0 versus 27.2%, respectively; P = 0.027). For old injury patients, the lunate necrosis rate was higher in those treated with the palmar approach than in those treated with the dorsal approach (50 versus 14.3%, respectively; P = 0.033). CONCLUSIONS: The key points for resolving distal radius fracture with lunate dislocation are prompt and precise diagnosis and treatment of lunate dislocation to prevent old lunate dislocation. We recommend that the surgical procedure is performed through the dorsal approach with reparation of the dorsal radiolunate ligament.

Current advances in animal model of chondrosarcoma and related research.

Chondrosarcoma is a type of malignant cartilage tumor with a high local recurrence. Due to its resistance to chemo- and radiotherapy, current treatment is limited to surgical resection. Animal model is one of the most important approaches to studying this disease, although systematic reporting on its development is rare. In this review, we summarized the elements involving animal model establishment. On the basis of these elements, we further classified chondrosarcoma animal models into various types. In addition, we compared various measurements for evaluating the animal model. Finally, its specific applications were discussed.

Modified hemiarthroplasty for four-part fractures of the proximal humerus.

BACKGROUND: Hemiarthroplasty is used to treat patients with Neer four-part fractures but the post-operative complication rate remains high, particularly with regard to shoulder joint instability. This study was aimed at evaluating the efficacy of a modified rotator cuff reconstruction and balance technique by comparing it with conventional rotator cuff restoration surgery for Neer four-part fractures. METHODS: This retrospective therapeutic study included 67 patients with Neer four-part fractures. Forty-five patients underwent the modified surgery and 22 control patients underwent conventional surgery. All patients were followed up for more than 3 years. Outcome was assessed by grading clinical outcome and pain and strength, measuring range of motion, and radiographic analysis. RESULTS: The Neer score was higher in the modified surgery group (84 versus 75; P = 0.009). The rate of satisfaction (Neer score ≥80) in the modified surgery group was higher than in the conventional surgery group (71.11% versus 40.91%; P = 0.017). Range of motion for active abduction, external rotation, internal rotation and active forward elevation was greater in the modified surgery group (all P < 0.01). The morbidity rate of post-operative instability of the shoulder joint in the modified surgery group was much lower than that in the conventional surgery group (2.22% versus 18.2%; P < 0.01). CONCLUSION: The modified hemiarthroplasty procedure for treatment of Neer four-part fractures prevents post-operative instability of the shoulder joint and improves function of the shoulder more effectively than the conventional hemiarthroplasty procedure.

The techniques of soft tissue release and true socket reconstruction in total hip arthroplasty for patients with severe developmental dysplasia of the hip.

PURPOSE: Total hip arthroplasty (THA) is an effective procedure for developmental dysplasia of the hip (DDH); however, it is sometimes difficult to complete for severe cases because of femoral head dislocation, dysplasia of the acetabulum and the femur, disparity in limb length, soft tissue contraction, and muscular atrophy. We aimed at exploring the efficiency of the techniques of release and balance of soft tissues and reconstruction of true socket THA for patients with severe DDH. METHODS: From January 2000 to January 2009, 46 adult patients with severe DDH (50 hips) were included in this study. According to the classification system, there were 26 type III and 24 type IV. Among them there were 32 women and 14 men, aged from 38 to 77 years. THA was performed via a lateral approach. All acetabular sockets were reconstructed at the original anatomical location following a meticulous technique of soft tissue release and balance around the hip to restore limb length, to strengthen the abductor and improve its function. RESULTS: All patients had restoration of limb length (range, 2.5-5.5 cm; 30 limbs of more than 4 cm) without injury to the sciatic nerve. One postoperative dislocation occurred due to slight enlargement of the angle of abduction of the acetabulum. The follow-up ranged from 2.2 to 11.5 years (median 6.4 years) in 46 patients, and the Harris score increased from 40.2 preoperatively to 86.5 (P = 0.027). All hips were pain free with good function at the latest follow-up. CONCLUSION: The meticulous techniques of soft tissue release and balance can be recommended to ensure anatomical reconstruction of the true acetabular socket and to improve abductor function during arthroplasty for DDH.

[Application of an integrated omics analysis for the discovery of biomarkers for osteosarcoma].

OBJECTIVE: To apply an integrated omics analysis so as to discover potential biomarkers for an early diagnosis of osteosarcoma. METHODS: Gene chip and surface-enhanced laser desorption/ionization-time of flight-mass spectra (SELDI-TOF-MS) were employed to screen the marker genes and serum markers of osteosarcoma respectively. The associations of potential biomarkers from the SELDI data and microarray analysis were further inferred by Link-test to identify stable and specific diagnostic biomarkers. RESULTS: A total of 653 pre-biomarkers were found in osteosarcoma cells. And six differentially expressed protein peaks with strong statistical significances were detected by SELDI-TOF-MS in the patient's serum samples. Thirteen potential biomarkers for an early diagnosis of osteosarcoma were screened by Link-test. CONCLUSION: As an effective method of identifying osteosarcoma biomarkers, the integrated omics analysis may serve as a monitoring platform for an early diagnosis of osteosarcoma.

Arcus venosus dorsalis pedis graft to restore the superficial palmar arch in devascularized hands.

For complex amputation injuries and anatomical characteristic of the metacarpus, restoration to the superficial palmar arch in severe devascularized or amputated hands remains a daunting challenge. In this article, we treated a series of five cases using an arcus venosus dorsalis pedis (AVDP) graft to restore the superficial palmar arches in devascularized hands. All hands survived and satisfying function was gained. The AVDP graft may be a suitable option to restore the superficial palmar arch in devascularized hands.