Deep-penetration functionalized cuttlefish ink nanoparticles for combating wound infections with synergetic photothermal-immunologic therapy.

The challenge of wound infections post-surgery and open trauma caused by multidrug-resistant bacteria poses a constant threat to clinical treatment. As a promising antimicrobial treatment, photothermal therapy can effectively resolve the problem of drug resistance in conventional antibiotic antimicrobial therapy. Here, we report a deep-penetration functionalized cuttlefish ink nanoparticle (CINP) for photothermal and immunological therapy of wound infections. CINP is decorated with zwitterionic polymer (ZP, namely sulfobetaine methacrylate-methacrylate copolymer) to form CINP@ZP nanoparticles. Natural CINP is found to not only exhibit photothermal destruction of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli), but also trigger macrophages-related innate immunity and enhance their antibacterial functions. The ZP coating on the surface of CINP enables nanoparticles to penetrate into deeply infected wound environment. In addition, CINP@ZP is further integrated into the thermosensitive Pluronic F127 gel (CINP@ZP-F127). After in situ spraying gel, CINP@ZP-F127 is also documented notable antibacterial effects in mice wound models infected with MRSA and E. coli. Collectively, this approach combining of photothermal therapy with immunotherapy can promote delivery efficiency of nanoparticles to the deep foci of infective wounds, and effectively eliminate wound infections.

Stemness Subtypes and Scoring System Predict Prognosis and Efficacy of Immunotherapy in Soft Tissue Sarcoma.

Tumor stemness has been reported to play important roles in cancers. However, a comprehensive analysis of tumor stemness remains to be performed to investigate the specific mechanisms and practical values of stemness in soft tissue sarcomas (STS). Here, we applied machine learning to muti-omic data of patients from TCGA-SARC and GSE21050 cohorts to reveal important roles of stemness in STS. We demonstrated limited roles of existing mRNAsi in clinical application. Therefore, based on stemness-related signatures (SRSs), we identified three stemness subtypes with distinct stemness, immune, and metabolic characteristics using consensus clustering. The low-stemness subtype had better prognosis, activated innate and adaptive immunity (e.g., infiltrating B, DC, Th1, CD8+ T, activated NK, gamma delta T cells, and M1 macrophages), more enrichment of metabolic pathways, more sites with higher methylation level, higher gene mutations, CNA burdens, and immunogenicity indicators. Furthermore, the 16 SRS-based stemness prognostic index (SPi) was developed, and we found that low-SPi patients with low stemness had better prognosis and other characteristics similar to those in the low-stemness subtype. Besides, low-stemness subtype and low-SPi patients could benefit from immunotherapy. The predictive value of SPi in immunotherapy was more accurate after the addition of MSI into SPi. MSIlowSPilow patients might be more sensitive to immunotherapy. In conclusion, we highlighted mechanisms and practical values of the stemness in STS. We also recommended the combination of MSI and SPi which is a promising tool to predict prognosis and achieve precise treatments of immunotherapy in STS.

Hardware removal or preservation? Decision making based on a newly developed rating scale.

Hardware exposure following open reduction and internal fixation (ORIF) surgery is a tricky problem. It is always hard for surgeons to decide whether to keep or remove the hardware. In this study, a rating scale and corresponding clinical path is developed based on former published paper as well as our own experience. New admitted patients are first evaluated and scored once they enter the department. Based on the score they get, patients are assigned to different therapeutic schedule, i.e. (1) hardware preservation with pedicel flap transplantation, (2) debridement for further reevaluation and (3) hardware removal with external fixation. Satisfying clinical outcome is achieved that is characterized with high osseous consolidation rate and low complication rate. The result showed that this newly developed rating scale and the related therapeutic schedule could be an available tool to help surgeons to make decisions in the treatment of hardware exposure.

Downregulation of IDH2 exacerbates the malignant progression of osteosarcoma cells via increased NF-κB and MMP-9 activation.

Isocitrate dehydrogenase 2 (IDH2) is a mitochondrial NADP-dependent isocitrate dehydrogenase. It is considered to be a novel tumor suppressor in several types of tumors. However, the role and related mechanism of IDH2 in osteosarcoma remain unknown. The expression and significance of IDH2 were investigated by immunohistochemistry in formalin-fixed paraffin sections from 44 osteosarcoma patients. IDH2 was downregulated via lentiviral vector­mediated RNA interference (RNAi) in the Saos-2 and MG-63 human osteosarcoma cell lines. The effect of IDH2 downregulation on human osteosarcoma was studied in vitro by MTT, flow cytometry and invasion assays. Nuclear factor-κB (NF-κB) and matrix metalloproteinase-9 (MMP-9) assays were also used to study the likely molecular mechanism of IDH2 downregulation on the malignant progression of osteosarcoma cells. The results revealed that the expression of IDH2 was inversely correlated with pathological grade and metastasis in osteosarcoma. IDH2 downregulation promoted a pro-proliferative effect on the Saos-2 and MG-63 osteosarcoma cell lines. IDH2 downregulation accelerated cell cycle progression from S to G2/M phase. The pro-proliferative effect induced by IDH2 downregulation may be ascribed to increased NF-κB activity via IκBα phosphorylation. The invasive activity of osteosarcoma cells was also significantly promoted by IDH2 downregulation and may result from elevated MMP-9 activity. In conclusion, IDH2 downregulation may exacerbate malignant progression via increased NF-κB and MMP-9 activity and may implicate the potential biological importance of IDH2 targeting in osteosarcoma cells. Downregulation of IDH2 exacerbates the malignant progression of osteosarcoma cells via increased NF-κB and MMP-9 activation.

A comparison of primary and delayed wound closure in severe open tibial fractures initially treated with internal fixation and vacuum-assisted wound coverage: a case-controlled study.

The ideal timing of wound closure for open tibial fractures is debatable. This study aimed to compare outcomes of primary and delayed wound closure in severe open tibial fractures initially treated with internal fixation and vacuum-assisted wound coverage (VAC). Data of 80 patients with Gustilo-Anderson type IIIA and IIIB open tibial fractures treated with primary internal fixation, VAC, either primary wound closure (PWC) or delayed wound closure (DWC), and external fixation were reviewed retrospectively, and outcomes and complications compared. Patients were divided into three groups, including a PWC group (n = 27), DWC group (n = 22), and a control group (n = 31) that had received external fixation. Among all patients, the median age was 38 years (IRQ 32-47 years), and 67.5% were male. Injuries included 33 Gustilo-Anderson type IIIA and 47 type IIIB. Among injuries, 83% (66/80) were high-energy trauma, 63.8% were contaminated and median injury severity score (ISS) was 14 points. Significant differences were found between groups in fixation methods (p < 0.001). No significant differences were observed between groups in rates of deep infection, osteomyelitis, amputation and nonunion at 6 and 12 months (all p > 0.05), although all rates were markedly lower in the PWC group. The outcomes of PWC performed in conjunction with primary internal fixation and VAC for the treatment of Gustilo-Andersen type IIIA and IIIB open tibial fractures are similar to or better than those of DWC with primary internal fixation and VAC.

Absorbable implants versus metal implants for the treatment of ankle fractures: A meta-analysis.

This meta-analysis was performed to evaluate the efficiency and the safety of absorbable implants. Five major electronic databases (PubMed, Embase, Cochrane Library, SinoMed and Wanfang Data) were systematically searched for randomized controlled trials (RCTs) from their establishment to November 2012. Studies on absorbable implants and metal implants for ankle fractures were selected. The meta-analysis was performed using RevMan 5.1. Ten studies with 762 patients were included and analyzed. Compared with metal implants, absorbable implants used for the internal fixation of ankle fractures produce similar radiographic and functional outcomes (P= 0.52). Normally, removal of the internal fixation is unnecessary (P<0.0001) and the incidence of palpable implants is lower (P=0.02) for absorbable implants. No statistically significant difference was observed between the two groups with regard to foreign body reactions (P=0.07), infection (P= 0.69), osteoarthritis (P= 0.39), pain (P= 0.06), refracture (P=0.67), skin necrosis (P=0.99), deep vein thrombosis (P=0.21) and nerve injury (P=0.94). Absorbable implants used in ankle fractures rarely require reoperation and result in similar functional outcomes and complications compared with metal implants. These characteristics make them efficient and reasonably safe for the treatment of ankle fractures.

Chitosan/poly(vinyl alcohol) hydrogel combined with Ad-hTGF-ß1 transfected mesenchymal stem cells to repair rabbit articular cartilage defects.

The aim of this work is to explore the feasibility and therapeutic effect of repairing rabbit articular cartilage defects using thermo-sensitive chitosan/poly (vinyl alcohol) composite hydrogel engineered Ad-hTGF-ß1-transfected bone marrow mesenchymal stem cells. Rabbit's bone marrow stromal cells (BMSCs) were obtained and cultured in vitro and transfected with a well-constructed Ad-hTGF-ß1 vector, the cartilage phenotype of the transfected cells was tested by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Twenty-four New Zealand white rabbits with articular cartilage defects were randomly divided into four groups: group A was treated with CS/PVA gel and transfected BMSCs; group B received CS/PVA gel and un-transfected BMSCs; group C was treated with CS/PVA gel alone and group D was the untreated control group. Experimental animals of each group were killed at 16 weeks after operation. General observation, Masson's trichrome staining and collagen II immunohistological staining of the specimens were performed to evaluate the repair effect. The Wakitani scoring method was used to evaluate the repair effect. RT-PCR and Western blot confirmed that the hTGF-ß1 gene was expressed in BMSCs and triggered the expression of specific markers of cartilage differentiation such as aggrecan mRNA and Collagen II in BMSCs after transfection with Ad-hTGF-ß1. Sixteen weeks after operation, the defects in group A had smooth and flat surfaces, and the defects appeared to have completely healed, exhibiting almost the same color and texture as the surrounding cartilage. Masson's trichrome staining showed that the cell arrangement and density of regenerated cartilage tissue in group A was not significantly different from that of normal cartilage tissue. The immunohistochemical staining of Col II showed a strong expression in group A and weak expression in group B, but no expression in groups C and D. According to the Wakitani score, the difference between experimental group A and all of the other groups was statistically significant (P < 0.01). To conclude, as a thermosensitive and injectable scaffold material, CS/PVA gel engineered with BMSCs transfected with hTGF-ß1 can effectively repair rabbit articular cartilage defects.

The expression and significance of IDH1 and p53 in osteosarcoma.

BACKGROUND: To detect the expression of isocitrate dehydrogenase 1 (IDH1) and transformation-related protein 53 (p53) in osteosarcoma and analyze the correlation between them and the clinico-pathological features. METHODS: The expressions of IDH1 and p53 were detected in human osteosarcoma cell lines (MG-63 and U2OS) by immunocytochemistry, Real-time PCR and Western Blotting. The expressions of IDH1 and p53 in formalin-fixed paraffin-embedded tissue sections from 44 osteosarcoma patients were determined by immunohistochemistry, and the correlation between them and clinicopagthological features were analyzed. None of these patients received chemotherapy prior to surgery. RESULTS: IDH1 is detected in osteosarcoma cell lines and biopsies. IDH1 expresses higher in U2OS cells with wild type p53 than in MG-63 cells with mutation p53. IDH1 correlates with histological Rosen grade and metastasis negatively. P53 correlates with histological Rosen grade, metastasis and overall survival in clinical osteosarcoma biopsies. Osteosarcoma patients with High IDH1 expression have a very high p53 expression. CONCLUSION: IDH1 may correlate with p53 and be a candidate biomarker for osteosarcoma correlate with histological Rosen grade and metastasis.

[Treatment of osteosarcoma with TNF-related apoptosis-inducing ligand or in combination with doxorubicin in animal experiment].

OBJECTIVE: To examine the effect of TNF-related apoptosis-inducing ligand (TRAIL) and in combination with doxorubicin (ADM) to xenografted tumors in nude mice and to explore its potential mechanism. METHODS: MG-63 cells (5 x 10(6)/ml) were suspended in 0.2 ml RPMI-1640 and inoculated subcutaneously into the lower limb of nude mice. Treatment groups were given TRAIL of different concentration or combination of TRAIL and ADM intraperitoneally. Normal saline was administrated in the control group. Anti-tumor effects were estimated by tumor volumes. Serum alkaline phosphatase (ALP) was detected by ALP kits. Induction of apoptosis in xenografted tumors was confirmed by TUNEL (TdT-mediated dUTP nick end labeling) assay. Expression of Bax was detected by immunohistochemical assay. Expression of TRAIL receptors was detected by RT-PCR assay. RESULTS: Growth curve of tumors indicated that tumors carried by TRAIL-treated mice grew more slowly than that with normal saline and 2 microg TRAIL was more effective, Also tumors treated with combination of TRAIL and ADM grew more slowly than any other group. ALP activities of each group were moderately different but significance was not reached. TUNEL showed that there were more apoptotic cells in the combination group than any other group. Immunohistochemical assay showed that expression of Bax was up-regulated in the combination group. RT-PCR showed that expression of TRAIL-R2 mRNA was up-regulated in the combination group. CONCLUSION: TRAIL can induce an effective apoptosis of osteosarcoma cells in vivo in a dose-dependent fashion. ADM can enhance the effect of TRAIL-mediated apoptosis. And up-regulations of Bax and TRAIL-R2 may be the involved mechanism.