Search

Supplementary Data Legends - PDF file 57K, Figure legends for supplementary data

Purpose: Giant cell tumors of bone (GCTB) exhibit aggressive bone lytic behavior. Studies have shown that interleukin 17A (IL-17A) is involved pathologic bone resorption in various skeletal disorders. Thus, we have investigated the role of IL-17A in GCTBs.Experimental Design: We evaluated the progression of GCTBs using Campanacci grading and Enneking staging systems in 74 patients with GCTB. The expression of IL-17A and the IL-17A receptor A (IL-17RA) was assessed in GCTB tissues and in both multinucleated giant cells (MNGC) and stromal cells cultured in vitro using immunostaining and reverse transcription PCR (RT-PCR). The effects of IL-17A on the osteolytic activity of the MNGCs and the proliferation of the stromal cells were investigated using the “pit” formation and MTT assays, respectively. The effects of IL-17A on the expression of proosteolytic factors were examined in primary cultured MNGCs and stromal cells using RT-PCR, Western blotting, and gene expression microarrays.Results: In GCTBs, we detected abundant levels of IL-17A, which were associated with tumor extension and grade. IL-17A is predominantly produced by MNGCs, whereas IL-17RA is expressed by both MNGCs and stromal cells in GCTBs. In the MNGCs, the IL-17A increased the mRNA expression of IL-17A and proosteolytic enzymes, and also enhanced osteolytic ability. In the stromal cells, the IL-17A stimulated cellular proliferation and the expression of proosteolytic factors, including RANKL through myc and STAT3, respectively. In addition, IL-17A stimulated in vivo tumor growth and the extent of angiogenesis in GCTBs.Conclusion: IL-17A stimulates the progression of GCTBs and might represent a useful candidate marker for progression and as a therapeutic target for GCTBs. Clin Cancer Res; 19(17); 4697–705. ©2013 AACR.

Supplementary Data - PDF file 126K, Supplementary Figure S1. The isolation and culture of MNGCs and SCs from GCTB samples. A. After a quick centrifuge, the raw MNGCs were stained with Hoechst 33342 for 1h and sorted by a flow cytometric cell sorter. The MNGCs were huge and contained high levels of DNA and were thus clearly gated and sorted. A representative TRAP staining of the isolated MNGCs was shown. D. SCs were isolated and a representative microscopic morphology was shown. Supplementary Figure S2. Gene microarray and pathway analysis. A, A heat map of genes that are up-regulated over 1.5-fold by IL-17A in IL-17RA+SCs. The IL-17RA+SCs were treated with DMSO or 10 ng/ml IL-17A. After 48 hrs of IL-17A treatment, RNAs were isolated from cells and then subjected to gene microarray analysis.IL-17A pathway in IL-17RA-positive stromal cells. B, Software-assisted identification of pathways affected by IL-17A exposure, as revealed by Ingenuity Pathway Analysis. Differentially-expressed proteins showing a +1.5-fold change or greater were selected and entered into IPA software to illustrate potential interactions. Supplementary Figure S3. Myc and STAT3 inhibitor treatment suppressed IL-17A stimulated cell proliferation and RANKL expression in IL-17RA+SCs. A, The Myc inhibitor 10058-F4 treatment blocked IL-17A stimulated IL-17RA+ SCs proliferation. B, Treatment with the STAT3 inhibitor AG490 blocked IL-17A-induced RANKL mRNA expression in IL-17RA+SCs. The results are expressed as the mean values plus-minus SD. a,b,c,d data not sharing the same letter are significantly different from one another in each group (P < 0.05) by ANOVA and Duncan's multiple range test

Gastrointestinal stromal tumor (GIST) is one of the most common mesenchymal tumors of the gastrointestinal tract by activating mutations in the related receptor tyrosine kinases KIT and PDGFRA. Few...

Nicotine is the main addictive component in cigarettes that motivates dependence on tobacco use for smokers and makes it difficult to quit through regulating a variety of neurotransmitter release and receptor activations in the brain. Even though nicotine has an analgesic effect, clinical studies demonstrated that nicotine abstinence reduces pain threshold and increases pain sensitivity in smoking individuals. The demand for opioid analgesics in nicotine abstinent patients undergoing surgery has greatly increased, which results in many side effects, such as nausea, vomiting, and respiratory depression, etc. In addition, these side effects would hinder patients' physical and psychological recovery. Therefore, identifying the neural mechanism of the increase of pain sensitivity induced by nicotine abstinence and deriving a way to cope with the increased demand for postoperative analgesics would have enormous basic and clinical implications. In this review, we first discussed different experimental pain stimuli (e.g., cold, heat, and mechanical pain)-induced pain sensitivity changes after a period of nicotine dependence/abstinence from both animal and human studies. Then, we summarized the effects of the brain neurotransmitter release (e.g., serotonin, norepinephrine, endogenous opioids, dopamine, and γ-aminobutyric acid) and their corresponding receptor activation changes after nicotine abstinence on pain sensitivity. Finally, we discussed the limits in recent studies. We proposed that more attention should be paid to human studies, especially studies among chronic pain patients, and functional magnetic resonance imaging might be a useful tool to reveal the mechanisms of abstinence-induced pain sensitivity changes. Besides, considering the influence of duration of nicotine dependence/abstinence and gender on pain sensitivity, we proposed that the effects of nicotine abstinence and individual differences (e.g., duration of abstinence from smoking, chronic/acute abstinence, and gender) on abstinence-induced pain sensitivity should be fully considered in formulating pain treatment protocols. In summary, this paper could deepen our understanding of nicotine abstinence-induced pain sensitivity changes and its underlying neural mechanism, and could also provide effective scientific theories to guide clinical pain diagnosis and treatment, which has important clinical significance.

Objective. To compare the immediate and short term effectiveness of Shi’s Daoyin therapy (DT) rather than the Melbourne Protocol (MP) in terms of pain, mobility, and isometric strength of cervical muscles in nonacute nonspecific neck pain patients. Material and Methods. A total of 114 nonacute nonspecific neck pain patients aged 20~50 years were recruited and randomly assigned to be treated by either Shi’s DT or the MP. 56 cases and 54 cases received treatment for 3 weeks and were evaluated before and after intervention and at 3-week follow-up in Shi’s DT group and MP group, respectively. The outcome measures were Chinese version of the Neck Disability Index (NDI), cervical range of motion (ROM), maximal voluntary isometric force (MVIF), and pain intensity (Numeric Pain Rating Scale, NPRS). Results. All outcomes of both groups showed statistically significant improvements after the intervention and at 3-week follow-up (P < 0.05), while no statistically significant difference was found in NDI between groups. When followed up after 3 weeks, the ROM in axial rotation was significantly greater in the Shi’s DT group (P < 0.05), and the NPRS in the Shi’s DT group was significantly lower than the MP group (P < 0.05). At the end of the treatment period, the MVIF in lateral bending in the Shi’s DT group had a lower value (P = 0.044) than in the MP group, but there was no significant difference in flexion and extension between the two groups. Conclusions. Both Shi’s DT and MP groups demonstrated an obvious reduction in pain intensity and improvements in neck mobility after a short term follow-up period. The improvement of Shi’s DT in disability and pain during functional activities is generally similar to that of the MP for the treatment of nonacute nonspecific neck pain.

DNA damage in oocytes can cause infertility and birth defects. DNA double-strand breaks (DSBs) are highly deleterious and can substantially impair genome integrity. Homologous recombination (HR)-mediated DNA DSB repair plays dominant roles in safeguarding oocyte quantity and quality. However, little is known regarding the key players of the HR repair pathway in oocytes. Here, we identified oocyte-specific gene Ooep as a novel key component of the HR repair pathway in mouse oocytes. OOEP was required for efficient ataxia telangiectasia mutated (ATM) kinase activation and Rad51 recombinase (RAD51) focal accumulation at DNA DSBs. Ooep null oocytes were defective in DNA DSB repair and prone to apoptosis upon exogenous DNA damage insults. Moreover, Ooep null oocytes exhibited delayed meiotic maturation. Therefore, OOEP played roles in preserving oocyte quantity and quality by maintaining genome stability. Ooep expression decreased with the advance of maternal age, suggesting its involvement in maternal aging.

It has been reported that cyclin-dependent kinase like 3 (CDKL3) plays a crucial role in cell proliferation and migration in several cancers. However, the function of CDKL3 in triple-negative breast cancer (TNBC) is still unclear. In the present study, immunohistochemistry (IHC) was conducted to detect the CDKL3 expression. CCK-8, flow cytometry, Transwell assays, and mice xenograft models, were performed to explore the roles of CDKL3 on the proliferation and migration of TNBC in vitro and in vivo. Besides, protein chip analysis was used to screen the potential pathways, which was further confirmed by promoter activity assay, western blotting, and CCK-8 assay. Our findings reveal a high expression of CDKL3 in TNBC tissues, which is closely related to a poor prognosis of patients with TNBC. In TNBC cells, CDKL3 knockdown inhibits cell proliferation and migration, whereas CDKL3 overexpression has exactly the opposite effect. Consistently, CDKL3 knockdown induces cell apoptosis in vitro but suppresses tumor growth in vivo. Furthermore, CDKL3 knockdown increases p53 expression and reduces cell viability, and these effects are significantly weakened by the p53 inhibitor, PFT-α. In conclusion, the current study highlights that CDKL3 promotes TNBC progressions via regulating the p53 signaling pathway, suggesting that CDKL3 is a novel therapeutic target for TNBC treatment.

Objective. To evaluate the efficacy and safety of Tongning Gel (TNG) compared to placebo-controlled (PC) for knee osteoarthritis (KOA). Methods. A multicentre, randomized, double-blinded, parallel, placebo-controlled, clinical trial was performed in 576 patients (432 patients in the TNG group, 144 patients in the PC group), and 1 in the experimental group withdrew due to nonuse of drug. Patients were randomized to receive TNG or PC applied to knee skin at 3g per time, 2 times per day, which lasted for 3 weeks. The Western Ontario and McMaster Universities Arthritis Index (WOMAC) pain score was used to evaluate the primary efficacy of TNG and WOMAC stiffness and physical function and total scores were used to evaluate the secondary efficacy of TNG. All participants who received at least one dose of study drug were included in the safety analysis. This trial has been registered in Chinese Clinical Trial Registry (no. CTR20131276). Results. Primary efficiency outcome: there were significant differences in the decreased value of WOMAC pain score between two groups (P<0.05), and the decreased value of WOMAC pain score in the TNG group were better than those in the PC group (P<0.05). Secondary efficiency outcome: the WOMAC total score, WOMAC stiffness score, WOMAC physical function score, and the decrease of the above indexes of the two groups of patients after treatment were statistically significant (P<0.05), and the improvement of the above indexes in the TNG group was better than that of the PC group (P<0.05). Safety Evaluation. A total of 42 adverse events were reported by 29 patients: 25 adverse events reported by 16 patients (3.71%) in the experimental group and 17 adverse events were reported by 13 patients (9.03%) in the control group. And 8 adverse reactions were reported by 6 patients including 2 adverse reactions by 2 patients (0.46%) in the experimental group and 6 adverse reactions by 4 patients (2.78%) in the control group. Two cases of significant adverse events occurred in the experimental group. Both groups had one serious adverse event, respectively, which were not relevant to the intervention. Conclusion. These results of the trial demonstrate that TNG is superior to placebo in the treatment of patients with KOA, and TNG can improve other symptoms of KOA, such as stiffness and physical function. TNG is safe for the treatment of knee osteoarthritis as a whole.

Background Chronic low back pain is a common disease in clinic. The prevention and treatment of this disease requires a great deal of medical care resources, which is now the third medical economic burden in China. Although non-steroidal analgesics have been shown to be effective for chronic low back pain, considering the obvious side effects of these drugs, more and more patients are inclined to employ non-drug therapies. Typical examples are exercise therapy and cognitive-behavioral therapy (CBT). Exercise therapy of traditional Chinese medicine is an important part of non-drug therapy in China, such as sinew-strengthening exercise (SSE). Originating from traditional Chinese medicine exercise therapy, it has long been used for the prevention and treatment of chronic low back pain in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine (SGH), but it has not been validated by clinical trials. In this trial, we will seek to determine whether SSE is an effective and safe treatment option for persons with chronic nonspecific low back pain (CNLBP) by comparing it with CBT and explore a new method to CNLBP.Methods/Design In this trial, we will randomize 480 adults with nonspecific chronic back pain to CBT and SSE arms (240 per group). Participants in both groups will be followed for 52 weeks after randomization with home practice. Interviewers will assess outcomes 4, 8, 26 and 52 weeks postrandomization. The primary outcome will be Roland-Morris Disability Questionnaire (RMDQ) with 26-week follow-up being the primary endpoint.Discussion If SSE is found to be an effective treatment option for patients with chronic back pain, it will become a meritorious addition to the current limited treatment approaches available to patients with significant psychosocial contributors to their pain.

Cadherin-6 (CDH6) is aberrantly expressed in cancer and closely associated with tumor progression. However, the functions of CDH6 in human osteosarcoma and the molecular mechanisms underlying CDH6 in osteosarcoma oncogenesis remain poorly understood. In this work, we assessed the role of CDH6 in human osteosarcoma and identified that the expression of CDH6 was closely related with the overall survival and poor prognosis of osteosarcoma patients. MicroRNAs (miRNAs) have been implicated as important epigenetic regulators during the progression of osteosarcoma. Using dual-luciferase reporter assays, we showed that miR-223-3p suppresses CDH6 expression by directly binding to the 3′ UTR of CDH6. miR-223-3p overexpression significantly inhibited cell invasion, migration, growth, and proliferation by suppressing the CDH6 expression in vivo and in vitro. Besides, CDH6 overexpression in the miR-223-3p-transfected osteosarcoma cells effectively rescued the inhibition of cell invasion, migration, growth, and proliferation mediated by miR-223-3p. Additionally, Kaplan-Meier analysis suggests that the expression of miR-223-3p predicts favorable clinical outcomes for osteosarcoma patients. Moreover, the expression of miR-223-3p was downregulated in osteosarcoma patients and was negatively associated with the expression of CDH6. Collectively, these data highlight that miR-223-3p/CDH6 axis is an important novel pleiotropic regulator and could early predict the metastatic potential in human osteosarcoma treatments., Quanbo Ji et al. demonstrate that CDH6 is correlated with the overall survival and prognosis of osteosarcoma patients and identified as a novel direct target of miR-223-3p. miR-223-3p overexpression inhibited cell proliferation, invasion, and migration, indicating miR-223-3p and CDH6 could be novel prognostic biomarkers and therapeutic targets for treating osteosarcoma.

This study was designed to retrospectively analyze the survival and prognostic factors in Chinese osteosarcoma patients received neoadjuvant chemotherapy or/and surgery in a single institution. A total of 365 patients with pathological proved osteosarcoma undergoing neoadjuvant chemotherapy or/and surgery in a single institution between December 1999 and December 2012 were retrospectively analyzed for the demographic, tumor-related, and treatment-related variables, prognostic factors for survival rate and chemotherapy response. There were 231 males and 134 females (ratio, 1.72:1). The average age was 21±10years, with peak age between 10 and 20 years old (62%, 226/365). Of 365 patients, 319 (87.4%) suffered from primary tumor only, and 46 (12.6%) had metastases upon initial presentation. The overall 5-year survival rate was 65%. Upon univariate analysis, tumor site (femur 60.3%; other long bone 70.2%; trunk 33.6%; P=0.012), primary metastases (yes 36.7%; no 68.9%; P=0.000), tumor response to preoperative chemotherapy (good 89.8%; poor 47.5%; P=0.001) and recurrence/metastases after treatment (yes 36.2%; no 63.8%; P=0.000) were associated with higher 5-year survival rate. All factors except tumor site maintained their significance in multivariate testing. Male sex and nonconventional subtype of tumor were related to a higher likelihood of poor chemotherapy response.The absence of metastases at initial presentation, negative local recurrence or metastases after treatment, and tumor response to chemotherapy are of independent prognostic value in osteosarcoma. The overall prognostic factors and survival in Chinese patients are similar to those patients reported in western countries.

Ambient backscatter (AB) communication has emerged as an energy-preserving technology in Internet of Things (IoT). Specifically, an AB transmitter communicates with its receiver by backscattering ambient radio frequency (RF) signals instead of active information transmissions. When not operating in the communication mode, the AB transmitter can instead harvest the RF energy to replenish its battery, e.g., for supplying sensing energy consumptions. In this paper, we consider the optimal operating mode selection problem of the AB system that the transmitter decides either backscatter information or harvest energy based on the time-varying ambient RF signal strength. In particular, we aim to maximize the long-term average communication data rate given that an average energy-harvesting budget constraint is met. We formulate the optimization as a 0-1 knapsack problem and propose a dynamic programming (DP) based solution algorithm and a reduced-complexity greedy algorithm. Furthermore, we simulate under various setups, and show that the two proposed algorithms can achieve significant performance improvement over other representative benchmark methods.

The expression levels of p16 and nm23-H1 genes in soft tissue sarcoma (STS) were evaluated to investigate correlation of the expression levels with the incidence and prognosis of STS. Tumor tissues and para-carcinoma normal tissues were collected from 64 STS patients. The messenger ribonucleic acid (mRNA) expression levels of p16 and nm23-H1 in the tissues were detected via reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and the protein expression levels of p16 and nm23-H1 in tissues were detected using immunohistochemistry. Spearman's correlation analysis was used for the correlation between expression levels of p16 and nm23-H1 in STS tissues and the correlation between p16 and nm23-H1 mRNA and protein expression. Moreover, the correlation of p16 and nm23-H1 expression levels in tumor tissues with pathological parameters and prognosis of STS patients were analyzed combined with clinical data. Results of RT-qPCR showed that mRNA expression levels of p16 and nm23-H1 in tumor tissues of STS patients were significantly lower than those in para-carcinoma normal tissues (P

With rapid improvements in industrialization and urbanization, antibiotics are now extensively used to prevent and treat human and animal diseases and husbandry and aquaculture. Some research has been conducted to assess the environmental distribution and risk level of antibiotics, but their distribution remains largely uncharacterized. Thus, this study investigated the distribution and abundance of 39 antibiotics belonging to five groups, and their associated risks in surface water around Luoma Lake in the north of Jiangsu province, China. Nineteen antibiotics were detected, at a detection frequency (DF) ranging from 2.27% to 100%. The total antibiotics (ΣABs) concentrations ranged from 34.91 to 825.93 ng/L, with a median concentration of 195.45 ng/L. Among these antibiotics, chlortetracycline (DF: 100%; median: 172.02 ng/L) was the dominant antibiotic, accounting for a median percentage of 91.0% of ΣABs concentrations. Spearman rank correlation method found a significant correlation between clindamycin (DF: 72.7%; median: 2.01 ng/L) and lincomycin (DF: 79.5%; median: 4.58 ng/L). The ecological risk quotient (RQ) values for two out of 44 sampling sites were higher than 1, indicating a high risk; 11.4% of the RQ values fell between 0.1 and 1, indicating a medium risk. Moreover, roxithromycin was found to be the dominant contributor to the ecological risk, accounting for a median of 79.7% of ΣABs. However, the total non-carcinogenic (

The novel chloro(dimethyl)octylsilane (C8)-functionalized [CeO2//SiO2] Janus fibers (denoted CeO2//SiO2-C8) have been successfully prepared by electrospinning technique and subsequent modification. The CeO2//SiO2-C8 Janus fiber composed of the side-by-side CeO2 fiber and SiO2 fiber modified by C8 is endowed with separate affinity sites and hydrophobic chains. It has a relatively high specific surface area of 127.4 m2 g−1 with an average pore size of 4.1 nm. The potential of CeO2//SiO2-C8 Janus fibers for the selective capture and individual MS detection of low-abundance endogenous peptides and phosphopeptides from the tryptic protein digest and human serum real sample was explored and evaluated for the first time. The experimental results demonstrate that the CeO2//SiO2-C8 Janus fiber can selectively extract phosphopeptides in virtue of high selective affinity of CeO2 to phosphate groups and enrich low-abundance peptides on account of the strong hydrophobic interaction of C8 chains. Moreover, the enrichment performance of the CeO2//SiO2-C8 probe is superior to that of commercial products (e.g. TiO2 nanoparticles and ZipTipC18 pipette tips) available. Overall, the efficient enrichment capability of CeO2//SiO2-C8 Janus fibers makes them a prospective candidate for the capture of low-abundance endogenous peptides and phosphopeptides from complex biological samples for individual MS detection.

To obtain nitrogen-doped carbon dots (N-CDs) for sensing Fe3+ under acidic conditions, robust and highly fluorescent N-CDs were prepared by one-pot hydrothermal treatment of dopamine and ethylenediamine. The N-CDs have a relatively uniform size of approximately 4.9 nm and good water dispersibility. The N-CDs exhibit strong visible fluorescence (ca. 497 nm) and high stability at low pH (i.e. 2–3). The FTIR spectra and the XPS analyses indicated that primary amines and distinctive catechol groups exist on the surface of the N-CDs. The FTIR spectra further prove that the catechol groups on their surfaces can be oxidized to the quinone species by Fe3+, contributing to fluorescence response to Fe3+. Thus, the N-CDs as fluorescence probes allowed the detection of Fe3+ under acidic conditions, which has been verified in this work.

Background: Studies on the applications of bone transport using the Ilizarov method for osteosarcoma (OS) patients with surgical resection and neoadjuvant chemotherapy are rare. Methods: A retrospective analysis was conducted in 10 patients with limb OS receiving limb-salvage treatment by Ilizarov method from 2007 to 2012 in our hospital. The general information, treatment outcomes and follow-up data of the patients were collected. Results: The mean length of the transported fragment and the mean transport distance of the affected limb were both 14 cm. The mean time in the external fixator was 34.2 ± 11.2 months (16–47 months) and the mean external fixation index (EFI) was 75 days/cm. The mean follow-up time was 68.6 ± 26.6 months (37–103 months). Seven patients underwent additional operations to treat the postoperative complications, and the mean number of operation was 1.7 times. Only one patient underwent amputation due to tumor relapse and all patients survived without tumor. The limb-salvage rate was 90%. At the time of external fixator removal, the ASAMI-bone score was good in 66.7% of patients and the ASAMI-function score was fair in 66.7% of cases. The mean MSTS score was 18.6 ± 3.2 (n = 9). At 10 months after fixator removal, both the ASAMI-bone score and ASAMI-function score were both excellent in 80% and good in 20% cases, and the mean MSTS score was further improved to 27.2 ± 1.11 (n = 5). Conclusion: Bone transport using the Ilizarov method can achieve good therapeutic effectiveness in the limb-salvage treatment for OS patients with neoadjuvant chemotherapy as long as the complications can be timely recognized and well managed. Keywords: Osteosarcoma (OS), Ilizarov method, Limb salvage, Limb reconstruction, Neoadjuvant chemotherapy