Mechanical stretch preconditioned adipose-derived stem cells elicit polarization of anti-inflammatory M2-like macrophages and improve chronic wound healing.

Transplantation of adipose-derived stem cells (ASCs) is a promising option in the field of chronic wounds treatment. However, the effectiveness of ASCs therapies has been hampered by highly inflammatory environment in chronic wound areas. These problems could be partially circumvented using efficient approaches that boost the survival and anti-inflammatory capacity of transplanted ASCs. Here, by application of mechanical stretch (MS), we show that ASCs exhibits increased survival and immunoregulatory properties in vitro. MS triggers the secretion of macrophage colony stimulating factor (M-CSF) from ASCs, a chemokine that is linked to anti-inflammatory M2-like macrophages polarization. When the MS-ASCs were transplanted to chronic wounds, the wound area yields significantly faster closure rate and lower inflammatory mediators, largely due to macrophages polarization driven by transplanted MS-ASCs. Thus, our work shows that mechanical stretch can be harnessed to enhance ASCs transplantation efficiency in chronic wounds treatment.

Engineered Probiotic-Based Personalized Cancer Vaccine Potentiates Antitumor Immunity through Initiating Trained Immunity.

Cancer vaccines hold great potential for clinical cancer treatment by eliciting T cell-mediated immunity. However, the limited numbers of antigen-presenting cells (APCs) at the injection sites, the insufficient tumor antigen phagocytosis by APCs, and the presence of a strong tumor immunosuppressive microenvironment severely compromise the efficacy of cancer vaccines. Trained innate immunity may promote tumor antigen-specific adaptive immunity. Here, a personalized cancer vaccine is developed by engineering the inactivated probiotic Escherichia coli Nissle 1917 to load tumor antigens and ß-glucan, a trained immunity inducer. After subcutaneous injection, the cancer vaccine delivering model antigen OVA (BG/OVA@EcN) is highly accumulated and phagocytosed by macrophages at the injection sites to induce trained immunity. The trained macrophages may recruit dendritic cells (DCs) to facilitate BG/OVA@EcN phagocytosis and the subsequent DC maturation and T cell activation. In addition, BG/OVA@EcN remarkably enhances the circulating trained monocytes/macrophages, promoting differentiation into M1-like macrophages in tumor tissues. BG/OVA@EcN generates strong prophylactic and therapeutic efficacy to inhibit tumor growth by inducing potent adaptive antitumor immunity and long-term immune memory. Importantly, the cancer vaccine delivering autologous tumor antigens efficiently prevents postoperative tumor recurrence. This platform offers a facile translatable strategy to efficiently integrate trained immunity and adaptive immunity for personalized cancer immunotherapy.

Deterministic and Probabilistic Health Risk Assessment of Toxic Metals in the Daily Diets of Residents in Industrial Regions of Northern Ningxia, China.

This study was designed to investigate the toxic metal (aluminum (Al), arsenic (As), chromium (Cr), cadmium (Cd), copper (Cu), nickel (Ni), lead (Pb), and zinc (Zn)) concentrations in drinking water and different foodstuffs meat (pork, beef, and mutton), cereals (rice, flour, corn, millet), beans (cowpeas, tofu), potatoes (potato, sweet potato), solanaceous fruits (pepper, eggplant, bitter gourd, cucumber), vegetables (cabbage, cauliflower, spinach), and fruits (apples, watermelons, pears, grapes)) and then estimate the potential health risks of toxic metal consumption to local residents in industrial regions of northern Ningxia, China. As in drinking water, Cr in meat, Pb in cereals, Pb in beans, As and Pb in potatoes, Pb in solanaceous fruits, Cr and Ni in vegetables, and Ni and Pb in fruits were the most contaminated heavy metals in the corresponding food with over-standard rates of 16.7%, 12.5%, 5.1%, 60%, 50%, 50%, 38.2%, 44.4%, 44.4%, 31.8%, and 31.8%, respectively.The results of the deterministic assessment of health risks showed that the total noncarcinogenic risk value of dietary intake of toxic metals by the local population was 5.6106, indicating that toxic metals pose a high noncarcinogenic risk. The order of the non-carcinogenic risk is HIcereal (1.2104) > HIsolanaceous fruit (0.9134) > HIVegetables (0.8726) > HIFruit (0.8170) > HIMeat (0.7269) > HIDrinking water (0.6139) > HIBeans (0.2991) > HIPotatoes (0.1573). The total carcinogenic health risk from exposure to toxic metals through dietary intake was 9.98 × 10-4, indicating that the total cancer risk value of residents is beyond the acceptable range (10-4) under the current daily dietary exposure and implies a high risk of cancer. The order of the carcinogenic risk is RDrinking water (2.34 × 10-4) > RMeat (2.11 × 10-4) > Rsolanaceous fruit (1.89 × 10-4) > RFruit (1.88 × 10-4) > Rcereal (1.36 × 10-4) > RPotatoes (2.44 × 10-5) > RVegetables (1.51 × 10-5) > RBeans (0). The probabilistic assessment results showed that 98.83% of the population is exposed to severe noncarcinogenic risk and 87.02% is exposed to unacceptable carcinogenic risk. The sensitivity analysis showed that drinking water, local cereals, vegetables, and fruits were the major contributors to health risks. Our results indicated that the daily dietary exposure of residents in industrial regions of northern Ningxia poses a serious threat to human health, and it is suggested that relevant departments should strengthen monitoring and control of the current situation of toxic metal pollution in the environment and continue to pay attention and take measures to reduce the exposure of toxic metals in the diets of residents in this area.

Machine Learning Predict Survivals of Spinal and Pelvic Ewing's Sarcoma with the SEER Database.

STUDY DESIGN: Retrospective Cohort Study. OBJECTIVES: This study aimed to develop survival prediction models for spinal Ewing's sarcoma (EWS) based on machine learning (ML). METHODS: We extracted the SEER registry's clinical data of EWS diagnosed between 1975 and 2016. Three feature selection methods extracted clinical features. Four ML algorithms (Cox, random survival forest (RSF), CoxBoost, DeepCox) were trained to predict the overall survival (OS) and cancer-specific survival (CSS) of spinal EWS. The concordance index (C-index), integrated Brier score (IBS) and mean area under the curves (AUC) were used to assess the prediction performance of different ML models. The top initial ML models with best performance from each evaluation index (C-index, IBS and mean AUC) were finally stacked to ensemble models which were compared with the traditional TNM stage model by 3-/5-/10-year Receiver Operating Characteristic (ROC) curves and Decision Curve Analysis (DCA). RESULTS: A total of 741 patients with spinal EWS were identified. C-index, IBS and mean AUC for the final ensemble ML model in predicting OS were .693/0.158/0.829 during independent testing, while .719/0.171/0.819 in predicting CSS. The ensemble ML model also achieved an AUC of .705/0.747/0.851 for predicting 3-/5-/10-year OS during independent testing, while .734/0.779/0.830 for predicting 3-/5-/10-year CSS, both of which outperformed the traditional TNM stage. DCA curves also showed the advantages of the ensemble models over the traditional TNM stage. CONCLUSION: ML was an effective and promising technique in predicting survival of spinal EWS, and the ensemble models were superior to the traditional TNM stage model.

Biodegradable electrospun nanofibrous platform integrating antiplatelet therapy-chemotherapy for preventing postoperative tumor recurrence and metastasis.

The perioperative trauma-related platelet recruitment and activation severely affect tumor recurrence and metastasis. Therefore, efficiently killing residual tumor cells and simultaneously inhibiting platelet activation to block platelet-cancer cell interaction might be a promising strategy to prevent postoperative tumor recurrence and metastasis. Methods: Biodegradable PLGA electrospun nanofibrous films co-delivering doxorubicin-loaded tumor repopulating cell-derived microparticles (DOX-MPs) and aspirin (ASA) were developed as the implant materials (DOX-MPs/ASA@NF) for postoperative in-situ treatment. The characterization, cytotoxicity against tumor cells, inhibition in platelet activation-triggered proliferation, migration and metastasis of tumor cells and in vivo anti-recurrence and anti-metastasis activity induced by DOX-MPs/ASA@NF were systematically evaluated. Results: PLGA nanofibrous films facilitate the enhanced distribution of DOX-MPs as well as DOX-MPs and ASA release in a time-programmed manner within the tumor resection cavity. The released DOX-MPs efficiently kill the residual tumor cells, while ASA decreases platelet activation and inhibits platelet-promoted proliferation, migration and metastasis of tumor cells, resulting in the remarkable inhibition of postoperative tumor recurrence and metastasis. Conclusions: DOX-MPs/ASA@NF may be a promising candidate to prevent the recurrence and metastasis of resectable tumors.

Reversing insufficient photothermal therapy-induced tumor relapse and metastasis by regulating cancer-associated fibroblasts.

Insufficient tumor accumulation and distribution of photosensitizers as well as low antitumor immunity severely restrict the therapeutic efficacy of photothermal therapy (PTT). Cancer-associated fibroblasts (CAFs) play a key role in tumor extracellular matrix (ECM) remodeling and immune evasion. Reshaping tumor microenvironment via CAF regulation might provide a potential approach for complete tumor elimination in combination with PTT. Here, tumor cell-derived microparticles co-delivering calcipotriol and Indocyanine green (Cal/ICG@MPs) are developed to modulate CAFs for improved PTT efficacy. Cal/ICG@MPs efficiently target tumor tissues and regulate CAFs to reduce tumor ECM, resulting in enhanced tumor accumulation and penetration of ICG to generate strong PTT efficacy and activate CD8+ T cell-mediated antitumor immunity. In addition, Cal/ICG@MPs-triggered CAF regulation enhances tumor infiltration of CD8+ T cells and ameliorates CAF-induced antigen-mediated activation-induced cell death of tumor-specific CD8+ T cells in response to PTT, eliciting long-term antitumor immune memory to inhibit tumor recurrence and metastasis. Our results support Cal/ICG@MPs as a promising drug to improve PTT efficacy in cancer treatment.

Levels and health risk assessment of pesticides and metals in Lycium barbarum L. from different sources in Ningxia, China.

The berries of Lycium barbarum L. (Goji) are widely used as a Chinese traditional herbal medicine and functional food because of their reported beneficial pharmacological effects. However, there are reports of Goji berries being contaminated by chemical residues that could pose a hazard to humans. In this study, samples of L. barbarum L. berries were collected from plantations in a genuine production area and supermarkets in Ningxia, China. The major hazardous chemicals, including pesticides (dichlorvos, omethoate, cypermethrin, fenvalerate, malathion, and deltamethrin) and metals (lead (Pb), cadmium (Cd), copper (Cu), nickel (Ni), zinc (Zn), and arsenic (As)), were quantified by gas chromatography and inductively coupled plasma-optical emission spectrometry. In addition, associated daily exposures and health risks were determined using deterministic and probabilistic assessments. The levels of five pesticides from the plantation samples were considerably lower than the maximum residue limits; only dichlorvos was detected in the supermarket samples, and deltamethrin was not detected in any samples. Cu, Zn, As, Pb, Ni and Cd were detected in samples from both sources. The hazard quotient values of individual hazardous chemicals and the hazard index of combined hazardous chemicals were considerably less than 1, indicating the absence of a non-carcinogenic effect of hazardous chemical exposures through Goji berry consumption. The R value of As was much less than 10-6, which shows that consumption of the Goji berries had no obvious carcinogenic risks. The potentially harmful effects of the L. barbarum L. are more likely from berries obtained from plantations than those from supermarkets, and metal exposure is more dangerous than pesticide exposure. However, on the basis of our analysis, no population would be exposed hazardous chemicals exceeding existing standards, and the factors most affecting the health risk were exposure frequency and As content.

Research topics and hotspot trends of lumbar spondylolisthesis: A text-mining study with machine learning.

Objectives: The study aimed to conduct a bibliometric analysis of publications concerning lumbar spondylolisthesis, as well as summarize its research topics and hotspot trends with machine-learning based text mining. Methods: The data were extracted from the Web of Science Core Collection (WoSCC) database and then analyzed in Rstudio1.3.1 and CiteSpace5.8. Annual publication production and the top-20 productive authors over time were obtained. Additionally, top-20 productive journals and top-20 influential journals were compared by spine-subspecialty or not. Similarly, top-20 productive countries/regions and top-20 influential countries/regions were compared by they were developed countries/regions or not. The collaborative relationship among countries and institutions were presented. The main topics of lumbar spondylolisthesis were classified by Latent Dirichlet allocation (LDA) analysis, and the hotspot trends were indicated by keywords with strongest citation bursts. Results: Up to 2021, a total number of 4,245 articles concerning lumbar spondylolisthesis were finally included for bibliometric analysis. Spine-subspecialty journals were found to be dominant in the productivity and the impact of the field, and SPINE, EUROPEAN SPINE JOURNAL and JOURNAL OF NEUROSURGERY-SPINE were the top-3 productive and the top-3 influential journals in this field. USA, Japan and China have contributed to over half of the publication productivity, but European countries seemed to publish more influential articles. It seemed that developed countries/regions tended to produce more articles and more influential articles, and international collaborations mainly occurred among USA, Europe and eastern Asia. Publications concerning surgical management was the major topic, followed by radiographic assessment and epidemiology for this field. Surgical management especially minimally invasive technique for lumbar spondylolisthesis were the recent hotspots over the past 5 years. Conclusions: The study successfully summarized the productivity and impact of different entities, which should benefit the journal selection and pursuit of international collaboration for researcher who were interested in the field of lumbar spondylolisthesis. Additionally, the current study may encourage more researchers joining in the field and somewhat inform their research direction in the future.

Pretreatment whole blood Epstein-Barr virus DNA predicts prognosis in Hodgkin lymphoma.

The study investigated pretreatment Epstein-Barr virus (EBV) DNA status and its prognostic values in 96 patients newly diagnosed Hodgkin lymphoma (HL). With 13.5 % patients in positive EBV DNA status before therapy, the positive group had inferior progression-free survival (PFS) (P = 0.023) as well as overall survival (OS) (P = 0.001). Pretreatment EBV DNA positivity was observed as an independent prognostic factor in OS (P = 0.036) while a trend to predict PFS (P = 0.064). By monitoring changes of EBV DNA copies in 13 patients with positive pretreatment EBV DNA status, 5 of 6 patients with complete response (CR) had their copies undetectable after 3 cycles of first-line treatment and 7 patients with progressive disease (PD) all had elevated EBV DNA copies during their relapsed period. Whole blood EBV DNA may be an adjunctive biomarker to reflect treatment response, risk of disease relapse as well as prognosis in HL patients.

25-Hydroxy vitamin D deficiency predicts inferior prognosis in Hodgkin lymphoma.

The study investigated serum 25-Hydroxy vitamin D (25-(OH)D) deficiency and its prognostic values of patients newly diagnosed Hodgkin lymphoma (HL). With seventy-seven patients enrolled, the median level of 25-(OH)D was 44.5 nmol/L (range, 15.5-100.9 nmol/L) and 16 (20.8 %) of them were considered as 25-(OH)D deficiency. With a median follow-up of 28 months (range, 4-56 months), the 2-year progression-free survival (PFS) and overall survival (OS) rate were 75.3 %±5.5 % and 94.7 %±3.0 %, respectively. Patients with deficient 25-(OH)D level had inferior PFS (P<0.001) as well as OS (P<0.001). In multivariate Cox analysis, 25-(OH)D deficiency was observed as an independent prognostic factor for both PFS (hazard ratio (HR) 3.323, 95 % CI 1.527-7.229, P = 0.002) and OS (HR 5.819, 95 % CI 1.322-25.622, P = 0.020). Receiver-operator characteristic (ROC) curve showed International Prognostic Score (IPS) plus 25-(OH)D deficiency (IPS-D) predicted more accurately than IPS in PFS (AUC: 0.735 (95 % CI 0.622-0.829) vs. 0.701 (95 % CI 0.586-0.800), P = 0.033) and OS (AUC: 0.864 (95 % CI 0.767-0.932) vs. 0.825 (95 % CI 0.722-0.902), P = 0.028). All these findings suggest that serum 25-(OH)D level may be an adjunctive indicator to predict prognosis in HL patient.

Iron chelation promotes 5-aminolaevulinic acid-based photodynamic therapy against oral tongue squamous cell carcinoma.

BACKGROUND: Oral tongue squamous cell carcinoma (OTSCC) is the most common malignancy of the oral cavity. Photodynamic therapy (PDT) has become a clinically promising approach for early stage OTSCC treatment. 5-Aminolaevulinic acid (5-ALA) is a precursor of protoporphyrin IX (PpIX) and has been applied for PDT of cancer. However, the accumulated PpIX in 5-ALA-treated cancer cells will be further transformed into heme through ferrous iron insertion under ferrochelatase catalysis. Theoretically, iron chelation can enhance the intracellular accumulation of PpIX and thus promote 5-ALA-based PDT. Here, an iron chelator deferasirox (DFX) was used to investigate synergistic suppression effects of 5-ALA-based PDT and iron chelation on OTSCC. METHODS: In OTSCC SCC-25 cells, the enhancing effect of DFX on 5-ALA-mediated accumulation of PpIX was firstly assessed. After laser irradiation (635 nm, 200 mW/cm2 and 2 min), the synergistic cytotoxicity and apoptosis-inducing effect of 5-ALA and DFX were evaluated in SCC-25 cells, and the apoptosis mechanism was further investigated by monitoring the change of mitochondrial membrane potential and observing the subcellular localization of cytochrome c (Cyt c). In SCC-25 tumor-bearing mice, the synergistic suppression effects of 5-ALA-based PDT and DFX on tumor growth and tumor angiogenesis were investigated after laser irradiation on the tumor (635 nm, 150 mW/cm2 and 10 min). RESULTS: In SCC-25 cells, DFX showed strong iron chelation effect and enhanced 5-ALA-mediated intracellular accumulation of PpIX by 2-3 folds. After laser irradiation (635 nm, 200 mW/cm2 and 2 min), 5-ALA combined with DFX exhibited significant synergistic effects on cytotoxicity and cell apoptosis. In the treated cells, the damage of mitochondrial membrane and the release of Cyt c from mitochondria to cytoplasm were observed distinctly, indicating the activation of mitochondria-related signal pathway. In SCC-25 tumor-bearing mice, tumor growth and tumor angiogenesis were both notably suppressed by combination treatment of 5-ALA with laser irradiation and DFX. Meanwhile, no obvious toxic injuries were visible in histological examination of major organs in the treated mice. CONCLUSIONS: 5-ALA-based PDT combined with iron chelation synergistically inhibited the growth of OTSCC. Hence it can be seen that this combination therapy may represent a promising strategy for clinical treatment of OTSCC and other cancers.

Glutamine-ß-cyclodextrin for targeted doxorubicin delivery to triple-negative breast cancer tumors via the transporter ASCT2.

Chemotherapy is the primary therapy for triple-negative breast cancer (TNBC) and the tumor-targeted delivery of chemotherapeutic drugs is necessary to minimize their side effects on normal tissues. TNBC cells display addictions to glutamine in culture, and the levels of the glutamine transporter, alanine-serine-cysteine transporter 2 (ASCT2), are elevated in many types of cancer. However, glutamine- or ASCT2-based carriers have not been used in tumor-targeted drug delivery. In this study, a novel derivative of ß-cyclodextrin (ß-CD), glutamine-ß-cyclodextrin (GLN-CD), was developed by conjugating glutamine with the 6-hydroxy of ß-CD, and GLN-CD was then used to prepare doxorubicin (DOX) inclusion complexes (DOX@GLN-CD) for TNBC treatment. GLN-CD and glutamine have similar ASCT2-binding sites, and GLN-CD has the potential to enter cells through ASCT2-dependent facilitated diffusion. An increase in the degree of substitution did not promote binding between GLN-CD and ASCT2. GLN-CD and DOX formed inclusion complexes at a molar ratio of 1 : 1. DOX@GLN-CD specifically accumulated in TNBC cells, including MDA-MB-231 and BT549 cells, where it subsequently induced G2/M blockade and apoptosis, but hardly affected nontumorigenic MCF10A cells. l-γ-Glutamyl-p-nitroanilide (GPNA), which is a specific inhibitor of ASCT2, antagonistically decreased the cellular uptake of DOX@GLN-CD by TNBC cells, which further confirmed the role of ASCT2 in DOX@GLN-CD transport. In vivo, DOX@GLN-CD accumulated specifically in tumors, achieved improved outcomes and minimized the toxic effects on main organs at the same dose as DOX. As a novel derivative of ß-CD, GLN-CD is an effective carrier that can specifically deliver DOX to TNBC cells via targeting ASCT2 and minimize its uptake by normal cells.

Multifunctional nanoparticles based on a polymeric copper chelator for combination treatment of metastatic breast cancer.

Copper plays an important role in tumor growth and metastasis. Copper chelation has been confirmed to be an effective strategy for breast cancer therapy through antiangiogenesis. In this work, a copper chelating coil-comb block copolymer RGD-PEG-b-PGA-g-(TETA-DTC-PHis) (RPTDH) was synthesized and used to prepare nanoparticles for loading resiquimod (R848), a TLR7 and TLR8 agonist, thus to combine antiangiogenesis and immune activation to treat breast cancer. RPTDH has strong copper-chelating ability and could self-assemble to form spherical nanoparticles with significant pH-sensitivity. R848 was efficiently loaded into RPTDH nanoparticles and exhibited greatly accelerated releases in weakly acid media simulating tumor microenvironment. RPTDH/R848 nanoparticles significantly inhibited the mobility, invasion and vascular tube formation of HUVECs via copper chelation, demonstrating their strong antiangiogenic activity in vitro. Furthermore, RPTDH/R848 nanoparticles remarkably induced the maturation and activation of human plasmacytoid dendritic CAL-1 cells, indicating their immune-activation ability. In breast tumor-bearing mice, RPTDH/R848 nanoparticles displayed excellent targeting ability for both primary breast tumor and lung metastases, and furthermore dramatically suppressed tumor growth and metastasis through copper deficiency-triggered antiangiogenesis and R848-induced immune activation. In summary, RPTDH/R848 nanoparticles can be used as an therapeutic agent against metastatic breast cancer through combining antiangiogenesis and immune activation.

Combination therapy of PKCζ and COX-2 inhibitors synergistically suppress melanoma metastasis.

BACKGROUND: Metastatic malignant melanoma is one of the most aggressive malignancies and its treatment remains challenging. Recent studies demonstrate that the melanoma metastasis has correlations with the heightened activations of protein kinase C ζ (PKCζ) and cyclooxygenase-2 (COX-2) signaling pathways. Targeted inhibitions for PKCζ and COX-2 have been considered as the promising strategies for the treatment of melanoma metastasis. Thus, the PKCζ inhibitor J-4 and COX-2 inhibitor Celecoxib were combined to treat melanoma metastasis in this study. METHODS: The Transwell assay, Wound-healing assay and Adhesion assay were used to evaluate the inhibition of combined therapy of J-4 and Celecoxib on melanoma cells invasion, migration and adhesion in vitro, respectively. The impaired actin polymerization was observed by confocal microscope and inactivated signal pathways about PKCζ and COX-2 were confirmed by the Western blotting assay. The B16-F10/C57BL mouse melanoma model was used to test the inhibition of combined therapy of J-4 and Celecoxib on melanoma metastasis in vivo. RESULTS: The in vitro results showed that the combination of J-4 and Celecoxib exerted synergistic inhibitory effects on the migration, invasion and adhesion of melanoma B16-F10 and A375 cells with combination index less than 1. The actin polymerization and phosphorylation of Cofilin required in cell migration were severely impaired, which is due to the inactivation of PKCζ related signal pathways and the decrease of COX-2. The combined inhibition of PKCζ and COX-2 induced Mesenchymal-Epithelial Transition (MET) in melanoma cells with the expression of E-Cadherin increasing and Vimentin decreasing. The secretion of MMP-2/MMP-9 also significantly decreased after the combination treatment. In C57BL/6 mice intravenously injected with B16-F10 cells (5 × 104 cells/mouse), co-treatment of J-4 and Celecoxib also severely suppressed melanoma lung metastasis. The body weight monitoring and HE staining results indicated the low toxicity of the combination therapy. CONCLUSIONS: This study demonstrates that the combination therapy of PKCζ and COX-2 inhibitors can significantly inhibit melanoma metastasis in vitro and in vivo, which will be an efficient strategy for treatment of melanoma metastasis in clinics.