Bacterial outer membrane vesicle-cancer cell hybrid membrane-coated nanoparticles for sonodynamic therapy in the treatment of breast cancer bone metastasis.

Breast cancer bone metastasis is a terminal-stage disease and is typically treated with radiotherapy and chemotherapy, which causes severe side effects and limited effectiveness. To improve this, Sonodynamic therapy may be a more safe and effective approach in the future. Bacterial outer membrane vesicles (OMV) have excellent immune-regulating properties, including modulating macrophage polarization, promoting DC cell maturation, and enhancing anti-tumor effects. Combining OMV with Sonodynamic therapy can result in synergetic anti-tumor effects. Therefore, we constructed multifunctional nanoparticles for treating breast cancer bone metastasis. We fused breast cancer cell membranes and bacterial outer membrane vesicles to form a hybrid membrane (HM) and then encapsulated IR780-loaded PLGA with HM to produce the nanoparticles, IR780@PLGA@HM, which had tumor targeting, immune regulating, and Sonodynamic abilities. Experiments showed that the IR780@PLGA@HM nanoparticles had good biocompatibility, effectively targeted to 4T1 tumors, promoted macrophage type I polarization and DC cells activation, strengthened anti-tumor inflammatory factors expression, and presented the ability to effectively kill tumors both in vitro and in vivo, which showed a promising therapeutic effect on breast cancer bone metastasis. Therefore, the nanoparticles we constructed provided a new strategy for effectively treating breast cancer bone metastasis.

Ozonated Hydrotherapy Combined with LED Yellow Light Irradiation and Oral Minocycline Treatment for Mild to Moderate Papulopustular Rosacea: A Comparative Retrospective Study.

Objective and Design: The treatment of recurrent rosacea has always been a problem. Oral minocycline has been widely used in the treatment of rosacea. However, the efficacy and safety of ozonated hydrotherapy combined with LED yellow light irradiation and oral minocycline for mild to moderate papulopustular rosacea (PPR) has not been thoroughly studied. Methods: Patients with rosacea who met the criteria and had complete clinical statistic admitted to our hospital from April 2021 to September 2022 were retrospectively collected and divided into combined therapy group and oral only group. The patients in the two groups were treated with minocycline for 8 weeks. In addition, the patients in combined therapy group were treated with ozone hydrotherapy once a week, followed by LED yellow light irradiation for a total of 4 weeks. The Investigator' s global assessment (IGA) score was used to assess the condition. The efficacy was evaluated using the patients' subjective symptom scores. Skin lesion images and adverse reactions were recorded. The recurrence rate was observed after 24 weeks of follow-up. Results: A total of 39 patients included in the study. After 4 weeks of treatment, the effective rate was 90% in combined therapy group and 52.63% in oral only group (p<0.05). After 8 weeks of treatment, the total score of the patients' subjective symptom scores and the scores of itching and burning sensation in combined therapy group were lower than those in oral only group (p<0.05). After 24 weeks of follow-up, the recurrence rate of combined therapy group was 5%, and that of oral only group was 26.32%. The mild adverse reactions experienced by both groups disappeared during follow-up. Conclusion: This combination therapy has a significant, rapid and safe therapeutic effect, especially in relieving itching and burning sensations, and may reduce the recurrence rate.

Engineered three-dimensional bioactive scaffold for enhanced bone regeneration through modulating transplanted adipose derived mesenchymal stem cell and stimulating angiogenesis.

Titanium alloy materials are commonly used in orthopedic clinical treatments. However, conventional titanium implants usually lead to insufficient bone regeneration and integration because of mismatched biomechanics and poor bioactivities. To tackle these challenges, a porous titanium alloy scaffold with suitable mechanical properties was prepared using three-dimensional (3D) printing, and then an adipose-derived mesenchymal stem cell (ADSC) loaded platelet-rich plasma (PRP) gel was placed into the pores of the porous scaffold to construct a bioactive scaffold with dual functions of enhancing angiogenesis and osteogenesis. This bioactive scaffold showed good biocompatibility and supported cell viability proliferation and morphology of encapsulated ADSCs. Osteogenic and angiogenic growth factors in the PRP gel promoted the migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) in vitro and enhanced osteogenic-related gene and protein expression in ADSCs, thus promoting osteogenic differentiation. After implantation into the femoral defects of rabbits, the bioactive scaffold promoted vascular network formation and the expression of osteogenesis-related proteins, thus effectively accelerating bone regeneration. Therefore, the osteogenic and angiogenic bioactive scaffold comprising a 3D printed porous titanium alloy scaffold, PRP, and ADSCs provides a promising design for orthopedic biomaterials with clinical transformation prospects and an effective strategy for bone defect treatment.

Factors Influencing Noise Following Primary Ceramic-on-Ceramic Total Hip Arthroplasty.

BACKGROUND: The noise associated with ceramic-on-ceramic (CoC) total hip arthroplasty (THA) has been a concerning issue, while its underlying causes remain unclear. METHODS: We conducted a retrospective analysis of 119 patients (174 primary CoC THAs) who had a mean follow-up of 28 months (range, 12 to 106). A questionnaire was designed to collect information on nature, frequency, onset, duration, and impact of the noise. Postoperative x-rays were evaluated. Clinical evaluations, including Harris and Oxford hip scores, were documented at follow-up time points (6 weeks, 3 months, 6 months, and 1 year). RESULTS: Of the 174 hips, 31.6% reported noise, including 26 popping (14.9%), 24 clicking (12.1%), and 5 grinding (2.9%). No patients reported squeaking. Noisy hips had lower age (P = .009) and body mass index (P = .019). Among patients with developmental dysplasia of the hip, 17 of 55 hips reported noise associated with smaller cup anteversion angle (P = .004), greater body height (P = .022), and larger acetabular cup size (P = .049). Noise typically began at a mean of 193 days (range, 1 to 2,598) after surgery and disappeared spontaneously in 50.9% of hips before final follow-up, with an average disappearance time of 211 days (range, 60 to 730). Noise did not affect daily life in 74.5% of patients, while 26.9% of patients who had popping reported painful sensations. One patient experienced joint dislocation, and another experienced a ceramic liner fracture during follow-up. No statistical difference was observed in outcome scores between noise and silent groups at 4 follow-up time points. CONCLUSIONS: Incidence of noise after primary CoC THA is relatively high. Smaller cup anteversion and larger acetabular cup size were associated with noise production in patients who had developmental dysplasia of the hip.

Machine learning-based estimation and mitigation of nitric oxide emissions from Chinese vegetable fields.

High fertilizer input and nitric oxide (NO) emissions characterize the intensive vegetable production system. However, the amount, geographic distribution, and effective mitigation strategies of NO emissions over Chinese vegetable fields remain largely uncertain. In this study, we developed a data-driven estimate of NO emissions and their spatial pattern in Chinese vegetable fields based on the Random Forest (RF) model. Additionally, we conducted a field experiment in a subtropical vegetable field to investigate the effect of climate-smart practices on NO emissions. The RF model results showed that soil NO emissions from Chinese vegetable fields were sensitive to nitrogen application amount, soil clay content, and pH. The total NO emission from Chinese vegetable fields in 2018 was estimated to be 75.9 Gg NO-N. The urgency to reduce NO emissions in vegetable fields was higher in northern than in southern China. Our meta-analysis and field experiment results suggested that biochar amendment and replacing chemical fertilizers with bio-organic fertilizers were win-win climate-smart management practices for mitigating NO emissions while improving vegetable production. Overall, our study provided new insights into NO emissions in vegetable soil ecosystems and can facilitate the development of regional NO emission inventories and effective mitigation strategies. These findings highlight the importance of adopting sustainable and climate-smart agricultural practices to reduce NO emissions and mitigate their adverse environmental impacts.

Lactate-upregulated NADPH-dependent NOX4 expression via HCAR1/PI3K pathway contributes to ROS-induced osteoarthritis chondrocyte damage.

Increasing evidence shows that metabolic factors are involved in the pathological process of osteoarthritis (OA). Lactate has been shown to contribute to the onset and progression of diseases. While whether lactate is involved in the pathogenesis of OA through impaired chondrocyte function and its mechanism remains unclear. This study confirmed that serum lactate levels were elevated in OA patients compared to healthy controls and were positively correlated with synovial fluid lactate levels, which were also correlated with fasting blood glucose, high-density lipoprotein, triglyceride. Lactate treatment could up-regulate expressions of the lactate receptor hydroxy-carboxylic acid receptor 1 (HCAR1) and lactate transporters in human chondrocytes. We demonstrated the dual role of lactate, which as a metabolite increased NADPH levels by shunting glucose metabolism to the pentose phosphate pathway, and as a signaling molecule up-regulated NADPH oxidase 4 (NOX4) via activating PI3K/Akt signaling pathway through receptor HCAR1. Particularly, lactate could promote reactive oxygen species (ROS) generation and chondrocyte damage, which was attenuated by pre-treatment with the NOX4 inhibitor GLX351322. We also confirmed that lactate could increase expression of catabolic enzymes (MMP-3/13, ADAMTS-4), reduce the synthesis of type II collagen, promote expression of inflammatory cytokines (IL-6, CCL-3/4), and induce cellular hypertrophy and aging in chondrocytes. Subsequently, we showed that chondrocyte damage mediated by lactate could be reversed by pre-treatment with N-Acetyl-l-cysteine (NAC, ROS scavenger). Finally, we further verified in vivo that intra-articular injection of lactate in Sprague Dawley (SD) rat models could damage cartilage and exacerbate the progression of OA models that could be countered by the NOX4 inhibitor GLX351322. Our study highlights the involvement of lactate as a metabolic factor in the OA process, providing a theoretical basis for potential metabolic therapies of OA in the future.

Near-Field Beamforming Algorithms for UAVs.

This study presents three distributed beamforming algorithms to address the challenges of positioning and signal phase errors in unmanned aerial vehicle (UAV) arrays that hinder effective beamforming. Firstly, the array's received signal phase error model was analyzed under near-field conditions. In the absence of navigation data, a beamforming algorithm based on the Extended Kalman Filter (EKF) was proposed. In cases where navigation data were available, Taylor expansion was utilized to simplify the model, the non-Gaussian noise of the compensated received signal phase was approximated to Gaussian noise, and the noise covariance matrix in the Kalman Filter (KF) was estimated. Then, a beamforming algorithm based on KF was developed. To further estimate the Gaussian noise distribution of the received signal phase, the noise covariance matrix was iteratively estimated using unscented transformation (UT), and here, a beamforming algorithm based on the Unscented Kalman Filter (UKF) was proposed. The proposed algorithms were validated through simulations, illustrating their ability to suppress the malign effects of errors on near-field UAV array beamforming. This study provides a reference for the implementation of UAV array beamforming under varying conditions.

Identification of Potentially Functional Circular RNA/Long Noncoding RNA-MicroRNA-mRNA Regulatory Networks Associated with Vascular Injury in Type 2 Diabetes Mellitus by Integrated Microarray Analysis.

This research is aimed at figuring out the potential circular RNA (circRNA)/long noncoding RNA- (lncRNA-) microRNA- (miRNA-) mRNA regulatory networks associated with a vascular injury in type 2 diabetes mellitus (T2DM). Differentially expressed genes (DEGs) screened in T2DM-related expression datasets were intersected with genes associated with vascular injury in T2DM to obtain candidate DEGs, followed by the construction of an interaction network of DEGs. The upstream miRNAs of candidate genes were predicted by mirDIP, miRWalk, and DIANA TOOLS databases, and the upstream lncRNAs/circRNAs of miRNAs by DIANA-LncBase/circBank database, followed by the construction of circRNA/lncRNA-miRNA-mRNA regulatory networks. Peripheral blood was attained from T2DM patients with macroangiopathy for clinical validation of expression and correlation of key factors. Differential analysis screened 37 candidate DEGs correlated with vascular injury in T2DM. Besides, MAPK3 was a core gene associated with vascular injury in T2DM. Among the predicted upstream miRNAs of MAPK3, miR-4270, miR-92a-2-5p, miR-423-5p, and miR-613 ranked at the top according to binding scores. The upstream lncRNAs and circRNAs of the 4 miRNAs were further predicted, obtaining 11 candidate lncRNAs and 3 candidate circRNAs. Moreover, KCNQ1OT1, circ_0020316, and MAPK3 were upregulated, but miR-92a-2-5p was downregulated in the peripheral blood of T2DM patients with macroangiopathy. Mechanistically, KCNQ1OT1 and circ_0020316 bound to miR-92a-2-5p that inversely targeted MAPK3. Collectively, KCNQ1OT1/circ_0020316-miR-92a-2-5p-MAPK3 coexpression regulatory networks might promote vascular injury in T2DM.

Platelet-rich plasma-enhanced osseointegration of decellularized bone matrix in critical-size radial defects in rabbits.

BACKGROUND: Bone defects represent a common orthopedic condition. With its vast array of donor sources, xenogeneic bone shows considerable potential as a bone defect repair material but may also be associated with immune rejection and reduced osteogenic capacity. Thus, reducing the risks for immune rejection of xenogeneic bone, while improving its osseointegration, are key technical challenges. METHODS: Decellularized bone matrix scaffolds (DBMs) were fabricated by thorough ultrasonic vibration and subjection to chemical biological agents to remove cells and proteins. The DBMs were then mixed with platelet-rich plasma (PRP) under negative pressure. Growth factor concentrations of PRP, as well as the microstructures and biomechanical properties of the system, were examined. Furthermore, osseointegration capacities in the critical-size radial defect rabbit model were verified. RESULTS: Complete decellularization of the scaffold and limited reductions in mechanical strength were observed. Moreover, the obtained PRP demonstrated various growth factors. Radiographic evaluation and histological analysis verified that more new bone formation occurred in the DBM mixed with PRP group at 6 and 12 weeks after implantation compared with both the blank group and the DBM without PRP group. CONCLUSIONS: Thorough physical and chemical treatments can reduce the probability of immune rejection of DBMs. The novel composite of DBMs mixed with PRP can serve as a promising bone regeneration material.

Endothelial cell-specific deficiency of the adenosine deaminase ADAR1 aggravates LPS-induced lung injury in mice via an MDA5-independent pathway.

Adenosine deaminase acting on RNA 1 (ADAR1) has been shown to participate in the regulation of endothelial cells (ECs), as well as local and systemic inflammatory responses. Here, we find that bacterial lipopolysaccharide (LPS)-induced upregulation of ADAR1 in lung ECs is impaired in aged mice, an animal model with high rates of sepsis and mortality. Endothelial cell-specific ADAR1 knockout (ADAR1ECKO ) mice suffer from higher mortality rates, aggravated lung injury, and increased vascular permeability under LPS challenge. In primary ADAR1 knockout ECs, expression of the melanoma differentiation-associated gene 5 (MDA5), a downstream effector of ADAR1, is significantly elevated. MDA5 knockout completely rescues the postnatal offspring death of ADAR1ECKO mice. However, there is no reduction in mortality or apoptosis in lung cells of ADAR1ECKO /MDA5-/- mice challenged with LPS, indicating the involvement of an MDA5-independent mechanism in this process.

Bisphosphonate-Functionalized Scaffolds for Enhanced Bone Regeneration.

The local sustained release of bioactive substances are attracting increasing attention in bone tissue engineering, which is beneficial to bone tissue formation and helps to improve the bone ingrowth ability of a scaffold. Bisphosphonates (BPs), as a representative kind of osteoclast inhibitors, are proven to possess excellent osteogenic induction capability. Accordingly, various physical and chemical strategies are developed to functionalize bone tissue scaffolds with BPs to achieve controlled release profiles. Compared with traditional treatment modalities, local release of BPs from these composite scaffolds will contribute to continuous bone integration without the risk of many complications. This review explores the molecular mechanisms of BPs on bone metabolism and analyzes the appropriate concentrations of BPs that promote bone regeneration. The advanced BP loading strategies, implant modification technologies, and BP-loaded composite scaffolds based on different matrices are summarized. Finally, the latest advances and the future development of BP-modified scaffolds for enhanced bone regeneration are discussed. This article provides leading-edge design strategies of the BP-functionalized bone engineering scaffolds for improved bone repairability.

[Degradation Mechanism of Tetracycline Using Fe/Cu Oxides as Heterogeneous Activators of Peroxymonosulfate].

A green, highly efficient, and porous copper-ferrite heterogeneous catalyst (Fe-Cu-400) with good magnetism was synthesized via a coprecipitation method. The catalysts were characterized using XRD, BET, FESEM, and EDS. The performance of Fe-Cu-400 as a catalyst was evaluated by activating peroxymonosulfate (PMS) for degradation of tetracycline (TC) in aqueous solution and investigating the influence of several water parameters. The Fe-Cu-400/PMS system showed a greater TC degradation ability, and the degradation rate of TC was enhanced with an increase in the PMS concentration and the initial pH of the coupled Fe-Cu-400/PMS system. Anions including H2PO4-, HCO3-, and Cl- promoted TC degradation, whereas NO3- showed a low inhibitory influence. In addition, Fe-Cu-400 exhibited excellent reusability towards activating PMS for TC degradation after five runs of tests. Possible mechanisms of the activation of PMS by Fe-Cu-400 and the main reactive species were proposed based on radical identification tests and XPS analysis. Furthermore, a potential degradation pathway was proposed that included hydrolysis and sequential removal of N-methyl, hydroxyl, and amine functional groups according to the results of LC-MS and TOC detections.

l-Homocysteine-induced cathepsin V mediates the vascular endothelial inflammation in hyperhomocysteinaemia.

BACKGROUND AND PURPOSE: Vascular inflammation, including the expression of inflammatory cytokines in endothelial cells, plays a critical role in hyperhomocysteinaemia-associated vascular diseases. Cathepsin V, specifically expressed in humans, is involved in vascular diseases through its elastolytic and collagenolytic activities. The aim of this study was to determine the effects of cathepsin V on l-homocysteine-induced vascular inflammation. EXPERIMENTAL APPROACH: A high methionine diet-induced hyperhomocysteinaemic mouse model was used to assess cathepsin V expression and vascular inflammation. Cultures of HUVECs were challenged with l-homocysteine and the cathepsin L/V inhibitor SID to assess the pro-inflammatory effects of cathepsin V. Transfection and antisense techniques were utilized to investigate the effects of cathepsin V on the dual-specificity protein phosphatases (DUSPs) and MAPK pathways. KEY RESULTS: Cathepsin L (human cathepsin V homologous) was increased in the thoracic aorta endothelial cells of hyperhomocysteinaemic mice; l-homocysteine promoted cathepsin V expression in HUVECs. SID suppressed the activity of cathepsin V and reversed the up-regulation of inflammatory cytokines (IL-6, IL-8 and TNF-α), adhesion and chemotaxis of leukocytes and vascular inflammation induced by l-homocysteine in vivo and in vitro. Increased cathepsin V promoted the degradation of DUSP6 and DUSP7, phosphorylation and subsequent nuclear translocation of ERK1/2, phosphorylation of STAT1 and expression of IL-6, IL-8 and TNF-α. CONCLUSIONS AND IMPLICATIONS: This study has identified a novel mechanism, which shows that l-homocysteine-induced upregulation of cathepsin V mediates vascular endothelial inflammation under high homocysteine condition partly via ERK1/2 /STAT1 pathway. This mechanism could represent a potential therapeutic target in hyperaemia-associated vascular diseases. LINKED ARTICLES: This article is part of a themed section on Spotlight on Small Molecules in Cardiovascular Diseases. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.8/issuetoc.

Additive manufacturing technique-designed metallic porous implants for clinical application in orthopedics.

Traditional metallic scaffold prostheses, as vastly applied implants in clinical orthopedic operations, have achieved great success in rebuilding limb function. However, mismatch of bone defects and additional coating requirements limit the long-term survival of traditional prostheses. Recently, additive manufacturing (AM) has opened up unprecedented possibilities for producing complicated structures in prosthesis shapes and microporous surface designs of customized prostheses, which can solve the drawback of traditional prostheses mentioned above. This review presents the most commonly used metallic additive manufacturing techniques, the microporous structure design of metallic scaffolds, and novel applications of customized prostheses in the orthopedic field. Challenges and future perspectives on AM fabricated scaffolds are also summarized.

Cementless total hip arthroplasty in advanced tuberculosis of the hip.

PURPOSE: The use of total hip arthroplasty (THA) to treat advanced tuberculous arthritis, particularly during the active phase, is challenging. The aim of this study was to evaluate the efficacy of cementless THA for advanced hip joint tuberculosis. METHODS: This study reviewed 32 patients (mean age at surgery, 49.4 years [range, 24-79 years]) who underwent cementless THA between 2007 and 2012. All patients were diagnosed with advanced tuberculosis of the hip on the basis of clinical manifestations, radiographic findings, and histological examination. All procedures were performed by a single surgeon. Mean follow-up period was 4.1 years (range, 2-7 years). Thorough debridement of tuberculosis-infected tissues and antitubercular therapy were conducted intra-operatively. Clinical data, including visual analog scale (VAS) score, Harris hip score (HHS), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and complications, as well as radiologic data, including prosthesis subsidence and loosening, bone growth, and heterotopic ossification, were evaluated during follow-up. RESULT: Mean VAS decreased from 7.6 (range, 5-10) pre-operatively to 1.4 (range, 0-4) at final follow-up (P < 0.01). Mean HHS improved from 42.2 (range, 30-75) pre-operatively to 85.4 (range, 60-95) at final follow-up (P < 0.01). No signs of reactivation were detected. In all patients, ESR and CRP levels were within normal limits by a mean of three and four months, respectively, and radiologic results during follow-up indicated favourable prosthesis positioning and condition. CONCLUSION: Despite the state of tuberculosis, cementless THA was an effective treatment for advanced tuberculosis of the hip.

Involvement of increased endogenous asymmetric dimethylarginine in the hepatic endoplasmic reticulum stress of type 2 diabetic rats.

OBJECTIVE: Increasing evidence suggested that endoplasmic reticulum (ER) stress contributes to insulin resistance, which plays an important role in the development of type 2 diabetes mellitus (T2DM). Accumulation of endogenous nitric oxide synthase (NOS) inhibitor, asymmetric dimethylarginine (ADMA), is associated with insulin resistance, T2DM, and diabetic cardiovascular complications, although the mechanisms have not been elucidated. This study was to determine whether elevated endogenous ADMA is involved in hepatic ER stress of type 2 diabetic rats, verify their causal relationship, and elucidate the potential mechanism underlying ADMA induced ER stress in rat hepatocytes. METHODS: Immunoglobulin binding protein (Bip) transcription, eukaryotic initiation factor 2α kinase (eIF2α) phosphorylation, X box-binding protein-1 (XBP-1) mRNA splicing and C/EBP homologues protein (CHOP) expression were measured to reflect ER stress. Contents of ADMA and nitrite/nitrate as well as activities or expression of NOS and dimethylarginine dimethylaminohydrolase (DDAH) were detected to show the changes in DDAH/ADMA/NOS/NO pathway. The lipid peroxidation product malondialdehyde content and antioxidant enzyme superoxide dismutase activity were analyzed to evaluate oxidative stress. RESULTS: ER stress was provoked in the liver of type 2 diabetic rats, as expressed by increases of Bip transcription, eIF2α phosphorylation, XBP-1 splicing and CHOP expression, all of which were in parallel with the elevation of serum ADMA, suppression of NO generation, NOS and DDAH activities in the liver. Exposure of hepatocytes to ADMA or hydrogen peroxide also induced ER stress, which was associated with the inhibition of NO production and increase of oxidative stress. Treatment of hepatocytes with antioxidant pyrrolidine dithiocarbamate not only decreased ADMA-induced oxidative stress and inhibition of NO production but also reduced ADMA-triggered ER stress. CONCLUSIONS: These results indicate that increased endogenous ADMA contributes to hepatic ER stress in type 2 diabetic rats, and the mechanism underlying ADMA-induced ER stress may relate to oxidative stress via NOS uncoupling.

Identification of DW532 as a novel anti-tumor agent targeting both kinases and tubulin.

AIM: 7,8-Dihydroxy-4-(3-hydroxy-4-methoxyphenyl)-2H-chromen-2-one (DW532) is one of simplified analogues of hematoxylin that has shown broad-spectrum inhibition on tyrosine kinases and in vitro anti-cancer activities. The aim of this study was to identify DW532 as a agent targeting both kinases and tubulin, and to investigate its anti-cancer and anti-angiogenesis activities. METHODS: In vitro tyrosine kinases activity was examined with ELISA, and tyrosine kinases activity in cells was evaluated with Western blot analysis. Tubulin turbidity assay, surface plasmon resonance and immunofluorescence technique were used to characterize the tubulin inhibitory activity. Cell proliferation was examined with SRB assay, and cell apoptosis and cell cycle distribution were analyzed with Annexin-V/PI staining and flow cytometry. Tube formation, aortic ring and chick chorioallantoic membrane assays were used to evaluate the anti-angiogenesis efficacy. RESULTS: DW532 inhibited EGFR and VEGFR2 in vitro kinase activity (the IC50 values were 4.9 and 5.5 µmol/L, respectively), and suppressed their downstream signaling. DW532 dose-dependently inhibited tubulin polymerization via direct binding to tubulin, thus disrupting the mitotic spindle assembly and leading to abnormal cell division. In a panel of human cancer cells, DW532 (1 and 10 µmol/L) induced G2/M phase arrest and cell apoptosis, which subsequently resulted in cytotoxicity. Knockdown of BubR1 or Mps1, the two core proteins of the spindle assembly checkpoint dramatically decreased DW532-induced cell cycle arrest in MDA-MB-468 cells. Moreover, treatment with DW532 potently and dose-dependently suppressed angiogenesis in vitro and in vivo. CONCLUSION: DW532 is a dual inhibitor against tubulin and tyrosine kinases, and deserves further development as a novel anti-cancer agent.

Oncologic outcomes of patients with Gleason score 7 and tertiary Gleason pattern 5 after radical prostatectomy.

PURPOSE: We evaluated oncologic outcomes following radical prostatectomy (RP) in patients with a Gleason score (GS) of 7 with tertiary Gleason pattern 5 (TGP5). MATERIALS AND METHODS: We retrospectively reviewed the medical records of 310 patients who underwent RP from 2005 to 2010. Twenty-four patients who received neoadjuvant or adjuvant antiandrogen deprivation or radiation therapy were excluded. Just 239 (GS 6 to 8) of the remaining 286 patients were included in the study. Patients were classified into four groups: GS 6, GS 7 without TGP5, GS 7 with TGP5, and GS 8. We analyzed preoperative clinical factors, postoperative pathological outcomes, and biochemical recurrence (BCR). RESULTS: TGP5 in GS 7 was an independent predictor of primary Gleason pattern 4, tumor volume larger than 10%, positive surgical margin, and lymphovascular invasion. The presence of TGP5 in GS 7 was not associated with BCR-free survival. Subgroup analyses revealed that BCR-free survival did not differ significantly between patients with GS 7 with TGP5 and those with GS 8 (p=0.120). In addition, time to BCR in patients with a higher percentage of TGP5 was shorter than that in patients with a lower percentage of TGP5. TGP5 in GS 7 was not a significant predictive factor for BCR, whereas prostate-specific antigen density and a positive surgical margin were shown to be independent predictors of BCR. CONCLUSIONS: TGP5 in GS 7 was an independent predictor of unfavorable pathologic outcomes. The rate of BCR was similar in GS 7 disease with TGP5 and in GS 8 disease, even though TGP5 was not a significant predictive factor for BCR in Cox proportional hazards models.

Contribution of endogenous inhibitor of nitric oxide synthase to hepatic mitochondrial dysfunction in streptozotocin-induced diabetic rats.

AIMS: Mitochondrial dysfunction plays important roles in the development of diabetes. Elevated nitric oxide (NO) synthase inhibitor asymmetric dimethylarginine (ADMA) has been shown to be closely related to diabetes. But the relationship between them in diabetes has not been determined. This study was to explore the role of ADMA in hepatic mitochondrial dysfunction and its potential mechanisms in diabetic rats and hepatocytes. METHODS: Respiratory enzymes activities, mitochondrial transmembrane potential and ATP content were measured to evaluate mitochondrial function. The copy number ratio of mitochondrial gene to nuclear gene was used to represent mitochondrial biogenesis. The activity of superoxide dismutase and malondialdehyde content were detected to reflect oxidative stress. Furthermore, changes in ADMA and NO contents, uncoupling protein 2 (UCP2) and peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) transcriptions were determined. RESULTS: Elevated ADMA levels in serum of diabetic rats were found to be associated with hepatic mitochondrial dysfunction reflected by reductions of respiratory enzyme activities, mitochondrial membrane potential and ATP contents. Similar mitochondrial dysfunction also occurred in ADMA-treated hepatocytes. The mitochondrial dysfunction observed in diabetic rats or hepatocytes was accompanied with suppressions of mitochondrial biogenesis, PGC-1α transcription and NO synthesis as well as enhances of UCP 2 transcription and oxidative stress. These effects of ADMA could be attenuated by treatments with antioxidant or NO donor. CONCLUSIONS: These results indicate that elevated endogenous ADMA contributes to hepatic mitochondrial dysfunction in diabetic rats, and underlying mechanisms may be related to the suppression of mitochondrial biogenesis and mitochondrial uncoupling via inhibiting NO synthesis and enhancing oxidative stress.