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.

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.

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.

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.

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.

[Reason analysis and treatment of acetabular component initial instability after primary total hip arthroplasty].

OBJECTIVE: To analyze the main reasons of acetabular component initial instability after primary total hip arthroplasty (THA) and to discuss the prevention and management. METHODS: The clinical data were retrospectively analyzed from 19 patients undergoing revision for acetabular component initial instability after primary THA between January 2003 and June 2010. There were 11 males and 8 females, aged from 55 to 79 years (mean, 67.2 years). The locations were left hip in 9 cases and right hip in 10 cases. The cementless hip prosthesis was used in 12 cases and cement hip prosthesis in 7 cases. The revisions were performed at 3 weeks to 6 months after primary THA. The reasons of early failure were analyzed. Both the coverage rate of acetabulum-bone and the Harris hip score were compared between pre- and post-revision. RESULTS: The main reason of acetabular component initial instability after primary THA may be unsuitable treatment of acetabulum, improper selection of acetabular component, and incorrect place angle of acetabular component. Sciatic nerve palsy occurred in 1 case and recovered at 7 weeks after revision. Slight fracture of the acetabulum in 1 case and healed at 3 months after revision. All incisions healed by first intention. No infection, vessel injury, displacement of acetabular component, or deep vein thrombosis occurred. The patients were followed up 11-73 months (mean, 28 months). At last follow-up, no acetabular component instability was observed. The coverage rate of acetabulum-bone was increased from 67.9% +/- 5.5% before revision to 87.7% +/- 5.2% after revision, showing significant difference (t = 11.592, P = 0.003). The Harris hip score at last follow-up (84.4 +/- 4.6) was significantly higher than that at pre-revision (56.5 +/- 9.3) (t = 11.380, P = 0.005). CONCLUSION: Detailed surgical plan, proper choice of component, correct place angle and elaborative planning, and proficient surgical skill are necessary to achieve the initial stability of acetabular component in THA.

Primary Classic Kaposi's Sarcoma of the Penis in an HIV-Negative Patient.

Kaposi's sarcoma (KS) is a multifocal hemorrhagic sarcoma that occurs primarily on the extremities. KS limited to the penis is rare and a well-recognized manifestation of acquired immune deficiency syndrome (AIDS). However, KS confined to the penis is extraordinary in human immunodeficiency virus (HIV)-negative patients. We present the case of a 68-year-old man with a dark reddish ulcerated nodule on the penile skin, which was reported as a nodular stage of KS. We detected no evidence of immunosuppression or AIDS or systemic involvements in further evaluations. In his past medical history, the patient had undergone three transurethral resections of bladder tumors due to urothelial cell carcinoma since 2000 and total gastrectomy, splenectomy, and adjuvant fluorouracil/cisplatin chemotherapy for 7 months due to advanced gastric carcinoma in 2005. The patient was circumcised and has had no recurrence for 2 years.