DLL1 orchestrates CD8+ T cells to induce long-term vascular normalization and tumor regression.

The immunosuppressive and hypoxic tumor microenvironment (TME) remains a major obstacle to impede cancer immunotherapy. Here, we showed that elevated levels of Delta-like 1 (DLL1) in the breast and lung TME induced long-term tumor vascular normalization to alleviate tumor hypoxia and promoted the accumulation of interferon γ (IFN-γ)-expressing CD8+ T cells and the polarization of M1-like macrophages. Moreover, increased DLL1 levels in the TME sensitized anti-cytotoxic T lymphocyte-associated protein 4 (anti-CTLA4) treatment in its resistant tumors, resulting in tumor regression and prolonged survival. Mechanically, in vivo depletion of CD8+ T cells or host IFN-γ deficiency reversed tumor growth inhibition and abrogated DLL1-induced tumor vascular normalization without affecting DLL1-mediated macrophage polarization. Together, these results demonstrate that elevated DLL1 levels in the TME promote durable tumor vascular normalization in a CD8+ T cell- and IFN-γ-dependent manner and potentiate anti-CTLA4 therapy. Our findings unveil DLL1 as a potential target to persistently normalize the TME to facilitate cancer immunotherapy.

Dexmedetomidine provides type-specific tumour suppression without tumour-enhancing effects in syngeneic murine models.

BACKGROUND: Dexmedetomidine is a widely used anaesthetic adjuvant for cancer resection surgeries. However, recent reports suggest that it may promote tumour growth or metastasis, so it is essential to clarify its tumour-related effects. METHODS: Seven syngeneic murine tumour models were used to assess the impact of dexmedetomidine on primary tumour growth, spontaneous tumour metastasis, and surgical resection-associated metastasis. Cancer cell proliferation and apoptosis experiments, terminal deoxynucleotidyl transferase dUTP nick-end labelling assays, immune cell analysis, specific T-cell depletion experiments, and gene transcription analysis were conducted to identify the underlying mechanisms. RESULTS: Dexmedetomidine did not affect growth of EO771 or 4T1 breast tumours, LAP0297 or LLC lung tumours, MCA205 fibrosarcoma, or their spontaneous lung metastases. It did not promote lung metastasis after breast cancer resection. Dexmedetomidine significantly suppressed MCA38 and CT26 colorectal tumour growth (P<0.01) and promoted apoptosis in MCA38 tumour tissues (P<0.05) without affecting proliferation and apoptosis of MCA38 tumour cells in vitro, suggesting indirect anti-tumour effects. Dexmedetomidine increased the proportions of intratumour CD4+ T (P<0.01), CD8+ T (P<0.001), and natural killer cells (P<0.01), and it upregulated transcription of the cytotoxicity-related genes Infg, Tnfa, and Cxcl9 (P<0.05) in MCA38 tumours. Either CD8+ or CD4+ T-cell depletion reversed the anti-tumour effects of dexmedetomidine on MCA38 tumours (P<0.05). CONCLUSIONS: Dexmedetomidine conferred colorectal tumour-type specific suppression by modulation of tumour CD4+ and CD8+ T cells without tumour-enhancing effects.

Vortex coagulation treatment of oilfield wastewater.

In this study, the traditional coagulation process was optimized and a method of vortex coagulation for wastewater treatment was proposed for the first time. The process discards the sedimentation method used in traditional coagulation and uses agitation to bring flocs up. These flocs can then be stuck by the filter cotton fixed in wastewater, and the filter cotton and flocs can act as net capturing several times under the action of agitation. It is worth noting that the filter cotton and flocs can synergistically adsorb the suspended and organic pollutants from wastewater. The results showed that the chemical oxygen demand (COD) removal of vortex coagulation could reach 82%, which was 10% higher than that of the traditional coagulation under the same coagulant dosing. In addition, the flocs in conventional coagulation and vortex coagulation were analyzed by IR spectroscopy and XRD, which showed that the mixing process did not destroy the chemical structure of the flocs. Compared with the conventional method, this process does not require a sedimentation tank, which can avoid the investment of equipment construction in this area. The effects of coagulant dosage, pH, stirring speed, and stirring time on the wastewater treatment effects were explored in the experiment.

CTLA4 blockade promotes vessel normalization in breast tumors via the accumulation of eosinophils.

Immune checkpoint blockade (ICB) has shown long-term survival benefits, but only in a small fraction of cancer patients. Recent studies suggest that improved vessel perfusion by ICB positively correlates with its therapeutic outcomes. However, the underlying mechanism of such a process remains unclear. Here, we show that anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA4) treatment-induced tumor vessel normalization was accompanied by an increased infiltration of eosinophils into breast tumors. Eosinophil accumulation was positively correlated with the responsiveness of a breast tumor to anti-CTLA4 therapy. Depletion of eosinophils subsequently negated vessel normalization, reduced antitumor immunity and attenuated tumor growth inhibition by anti-CTLA4 therapy. Moreover, intratumoral accumulation of eosinophils relied on T lymphocytes and interferon γ production. Together, these results suggest that eosinophils partially mediate the antitumor effects of CTLA4 blockade through vascular remodeling. Our findings uncover an unidentified role of eosinophils in anti-CTLA4 therapy, providing a potential new target to improve ICB therapy and to predict its efficacy.

LncRNA MSC-AS1 promotes osteogenic differentiation and alleviates osteoporosis through sponging microRNA-140-5p to upregulate BMP2.

This study aims to explore the role of lncRNA MSC-AS1/microRNA-140-5p/BMP2 regulatory loop in promoting osteogenic differentiation of BMSCs. BMSCs were isolated from bone marrow of mice. Expression levels of MSC-AS1, microRNA-140-5p and BMP2 during osteogenic differentiation were detected by qRT-PCR. Meanwhile, regulatory effect of MSC-AS1 on osteogenic differentiation was detected through ALP staining and alizarin red staining. The binding sites between microRNA-140-5p and MSC-AS1 as well as between microRNA-140-5p and BMP2 were predicted by TargetScan, which were further confirmed by dual-luciferase reporter gene assay. In addition, protein levels of MSC-AS1/microRNA-140-5p/BMP2 were detected by Western blot. Finally, rescue experiments were conducted to clarify the regulatory effects of MSC-AS1/microRNA-140-5p/BMP2 axis on osteogenic differentiation. MSC-AS1 and BMP2 were found to be remarkably up-regulated during osteogenic differentiation, while microRNA-140-5p was conversely down-regulated. Meanwhile, knockdown of MSC-AS down-regulated expression levels of osteogenesis-associated genes and weakened the mineralization capacity of BMSCs. MicroRNA-140-5p was verified to bind to the 3'UTR of MSC-AS1 and BMP2 genes. Knockdown of MSC-AS1 in BMSCs could reduce the expression of microRNA-140-5p, while knockdown of microRNA-140-5p also down-regulated BMP2 level. In addition, co-silence of MSC-AS1 and microRNA-140-5p reversed the inhibitory effect of MSC-AS1 knockdown on osteogenic differentiation and protein levels of p-Smad1/5/8, RUNX2 and Osterix. MSC-AS1 might promote the osteogenic differentiation of BMSCs through sponging microRNA-140-5p to up-regulate BMP2, thus alleviating the progression of osteoporosis.

Ferrate (IV) synthesis using derived persulfate during treatment of oil recovery wastewater.

Using oil recovery wastewater as the target material, the degradation of organic matter in oilfield wastewater was studied in an anodic oxidation system using Ti/Ru-Ir oxide-coated anode and 0.7mMNa2SO4 as the electrolyte. The TOC of the wastewater was 210 mg/L at the beginning of the electrolysis in the electrolysis system, and it decreased from 210 to 93.6 mg/L after 50 min of electrolysis. Under the action of this system, sulfate was oxidized to persulfate, and the apparent concentration of persulfate was 15.02 mM, oxidation and degradation of organic matter in wastewater by the action of newly generated persulfate.. Afterwards, NaOH and Fe2(SO4)3 were added to the electrolyzed wastewater, and the TOC in the wastewater was further reduced to 25.1 mg/L due to the coagulation effect of the newly generated Fe(OH)3 precipitate. The TOC removal rate of the wastewater treated by this process reached 88.0%, which meets the discharge requirements. At the same time, the derived persulfate oxidized Fe(OH)3 to generate a green substance, which was identified as Na2FeO3 by IR, UV, and XRD analyses. Na2FeO3 served as a highly effective water-purifying agent, demonstrating superior performance when compared to FeO42-. The method reported in this study not only provides a strategy for waste resource recycling but also offers the potential for mass production of ferrate (IV).

Distribution and characteristics of microplastics in sediment at representative dredged material ocean dumping sites, China.

Dredged material ocean dumping activities are likely an important source of microplastics (MPs) in coastal areas but have received little attention globally. In this study, we investigated the spatiotemporal distribution and characteristics of MPs in sediments at eight dredged material dumping sites of China. MPs were separated from sediment through density flotation, and polymer types were identified using µ-FTIR. The results showed that the average MP abundance was 112.82 ± 109.68 items/kg d.w. The MPs were more abundant at nearshore dumping sites than at distant dumping sites. Dumping activities may be the main contributor of MPs to Site BD1, the farthest dumping site from shore, but only a minor source of MPs at the other dumping sites. The characteristics of MPs were dominated by transparent PET fibers <1 mm. Overall, sediments at the dumping sites exhibited relatively low to moderate concentrations of MPs in comparison to most other coastal sediments.

Alleviating hypoxia to improve cancer immunotherapy.

Tumor hypoxia resulting from abnormal and dysfunctional tumor vascular network poses a substantial obstacle to immunotherapy. In fact, hypoxia creates an immunosuppressive tumor microenvironment (TME) through promoting angiogenesis, metabolic reprogramming, extracellular matrix remodeling, epithelial-mesenchymal transition (EMT), p53 inactivation, and immune evasion. Vascular normalization, a strategy aimed at restoring the structure and function of tumor blood vessels, has been shown to improve oxygen delivery and reverse hypoxia-induced signaling pathways, thus alleviates hypoxia and potentiates cancer immunotherapy. In this review, we discuss the mechanisms of tumor tissue hypoxia and its impacts on immune cells and cancer immunotherapy, as well as the approaches to induce tumor vascular normalization. We also summarize the evidence supporting the use of vascular normalization in combination with cancer immunotherapy, and highlight the challenges and future directions of this overlooked important field. By targeting the fundamental problem of tumor hypoxia, vascular normalization proposes a promising strategy to enhance the efficacy of cancer immunotherapy and improve clinical outcomes for cancer patients.

Bortezomib-resistant multiple myeloma patient-derived xenograft is sensitive to anti-CD47 therapy.

Multiple myeloma (MM) remains an incurable hematologic malignancy due to its frequent drug resistance and relapse. Cluster of Differentiation 47 (CD47) is reported to be highly expressed on MM cells, suggesting that the blockade of CD47 signaling pathway could be a potential therapeutic candidate for MM. In this study, we developed a bortezomib-resistant myeloma patient-derived xenograft (PDX) from an extramedullary pleural effusion myeloma patient sample. Notably, anti-CD47 antibody treatments significantly inhibited tumor growth not only in MM cell line-derived models, including MM.1S and NCI-H929, but also in the bortezomib-resistant MM PDX model. Flow cytometric data showed that anti-CD47 therapy promoted the polarization of tumor-associated macrophages from an M2- to an M1-like phenotype. In addition, anti-CD47 therapy decreased the expression of pro-angiogenic factors, increased the expression of anti-angiogenic factors, and improved tumor vascular function, suggesting that anti-CD47 therapy induces tumor vascular normalization. Taken together, these data show that anti-CD47 antibody therapy reconditions the tumor immune microenvironment and inhibits the tumor growth of bortezomib-resistant myeloma PDX. Our findings suggest that CD47 is a potential new target to treat bortezomib-resistant MM.

Spectrophotometric method for superoxide anion radical detection in a visible light (400-780 nm) system.

A method was developed that used hydroxylamine hydrochloride as a probe in a superoxide anion radical-generating visible light system for determining superoxide anion radicals. An Azure I solution with hydroxylamine hydrochloride was illuminated, after which a ferric iron solution was added to the sample solution and the remaining hydroxylamine hydrochloride in solution reduced from ferric to ferrous ions. Then, 1,10-phenanthroline solution was added and spectrophotometrically measured at 510 nm, which indirectly indicated the hydroxylamine hydrochloride content. The yields of superoxide anion radicals were indirectly expressed by the hydroxylamine hydrochloride decrement. Under optimal experimental conditions, the linear range was 0.0-1.5 × 10-5 M and the limit of detection and limit of quantification were obtained to be 8.37 × 10-7 and 2.54 × 10-6 M with an R2 of 0.9993. The method was simple and feasible and could be used for the stable measurement of superoxide anion radicals produced by photosensitizers that produce color under acidic conditions in visible light systems.

lncRNA LINC01535 upregulates BMP2 expression levels to promote osteogenic differentiation via sponging miR­3619­5p.

Osteoporosis is a debilitating skeletal disease that causes bones to collapse and is accompanied by a high risk of bone fracture. It was previously demonstrated that the osteogenic differentiation of human bone marrow­derived mesenchymal stem cells (hBMSCs) serves an important role in the process of human bone formation. Accumulating research has indicated that long non­coding RNAs (lncRNAs) participate in hBMSC osteogenic differentiation. For example, LINC01535 was reported to serve as a carcinogenic factor in cervical cancer; however, its latent function and molecular mechanism in the osteogenesis of hBMSCs remain to be investigated. The present study showed that the expression levels of LINC01535 were upregulated upon increasing osteogenic differentiation time. In addition, the inhibition of LINC01535 inhibited hBMSC proliferation and osteogenic differentiation and promoted cell apoptosis. Using bioinformatics analysis, LINC01535 was discovered to have complementary binding sites for microRNA (miR)­3619­5p, and further experiments demonstrated that LINC01535 functioned as a sponge of miR­3619­5p. Additionally, bone morphogenetic protein 2 (BMP2) was confirmed to be a target of miR­3619­5p. The results revealed that LINC01535 regulated the expression levels of BMP2 via sponging miR­3619­5p. In conclusion, the findings of the present study suggested that LINC01535 may accelerate the osteogenic process of hBMSCs via targeting the miR­3619­5p/BMP2 axis, which may offer an innovative therapeutic method for osteoporosis.

Genetic estrogen receptor alpha gene PvuII polymorphism in susceptibility to knee osteoarthritis in a Chinese Han population: A southern Jiangsu study.

BACKGROUND: Knee osteoarthritis (KOA) is the most prevalent type of arthritis and genetic factors play an important role in KOA pathogenesis. Some studies have reported the association of estrogen receptor alpha (ESRα) gene polymorphism and KOA susceptibility in different populations. This study was designed to verify whether ESRα gene polymorphism (rs2234693) was associated with primary KOA in a Chinese Han population living in the south of Jiangsu. METHODS: A case-control association study on single nucleotide polymorphism (SNP) rs2234693 was performed, and a total of 1953 subjects (1033 OA cases and 920 controls) were genotyped. Allele and genotype frequencies were compared between KOA cases and control participants. RESULTS: SNP rs2234693 was significantly associated with KOA in the dominant genetic model (TT + TC vs. CC) in all the subjects (odds ratio (OR) = 1.30; 95% confidence interval (CI) = 1.02-1.66; P = .03), and T allele frequency was also higher compared with allele C (OR = 1.38; 95% CI = 1.06-1.80; P = .02). After stratification by gender, there was no evident difference between the two groups in female and male subjects (P > .05). With a stratification for KOA severity, the combined genotype (TT + TC) (OR = 1.47; 95% CI = 1.12-1.94; P < .01) and T allele (OR = 1.61; 95% CI = 1.19-2.19; P < .01) were evidently associated with mild KOA, but not with severe KOA. CONCLUSIONS: ESRα gene is of considerable importance in the pathogenesis of early-stage KOA in a Chinese Han population living in southern Jiangsu.

MiR-337-3p promotes chondrocytes proliferation and inhibits apoptosis by regulating PTEN/AKT axis in osteoarthritis.

BACKGROUND: Osteoarthritis (OA) is a degenerative disease of articular cartilage and its main pathological feature is cartilage destruction, but its specific pathogenesis is still debatable. The aim of this study was to explore the role of miR-337-3p in OA pathogenesis. METHODS: The expression of miR-337-3p and PTEN in osteoarthritic cartilage tissues was detected using quantitative real time PCR and western blot, respectively. The regulation of miR-337-3p on PTEN was examined by luciferase reporter gene assays. The manipulation of miR-337-3p and PTEN was mediated by siRNA interference technology. The cell viability was analyzed by MTT assays. RESULTS: MiR-337-3p expression was significantly down-regulated in osteoarthritic cartilage tissues compared with normal cartilage tissues. Further studies confirmed that miR-337-3p overexpression evidently promoted the proliferation and inhibited the apoptosis of OA chondrocytes. PTEN expression was significantly up-regulated in osteoarthritic cartilage tissues and was negatively regulated by miR-337-3p in chondrocytes. PTEN silencing could improve the proliferation of OA chondrocytes and increased pAKT protein expression in OA chondrocytes. CONCLUSION: MiR-337-3p regulated OA chondrocytes proliferation through PTEN/AKT axis and thus involved in OA.

[Using a two-stage SBR process for removal of organics and nitrification-denitrification via nitrite].

High efficiency removal of organics and nitrogen by using a two-stage SBR process was introduced in this paper. Most of organics was removed in the first stage SBR reactor(SBR1) under the aerobic condition. Subsequently the second stage SBR reactor(SBR2) firstly operated under the aerabic condition for simultaneous nitrification and removal of a small amount of residual organics. Nitrification was controlled to the nitrite-type nitrification. Then denitrification happened in SBR2 under the anoxic condition. The petrochemical industry wastewater was used as external carbon sources in the denitrification. The experimental results indicated that in a two-stage SBR system, two kinds of biomass with the different function existed in the different reactors, which was beneficial to improve the treatment efficiency. The effluent COD reduced again because SBR2 removed COD which was left in SBR1 effluent. It resisted the disturbance of the high organic loading to nitrification. Consequently, as compared to a single SBR process, a two-stage SBR not only improved the treatment efficiency, but also saved the energy cost.