Publications by authors named "Wen Ju"

68 Publications

Dimethyl fumarate inhibits antibody-induced platelet destruction in immune thrombocytopenia mouse.

Thromb J 2021 Aug 28;19(1):61. Epub 2021 Aug 28.

Blood Diseases Institute, Xuzhou Medical University, 84 West Huaihai Road, Quanshan District, Xuzhou, 221002, Jiangsu, China.

Background: Immune thrombocytopenia (ITP) is an autoimmune disease characterized as a low platelet count resulting from immune-mediated platelet destruction. Dimethyl fumarate (DMF) is widely applied for the treatment of several autoimmune diseases with immunosuppressive effect. However, whether it ameliorates ITP is unclear. This study aims to evaluate whether DMF has a preventive effect on ITP in mice.

Methods: DMF (30, 60 or 90 mg/kg body weight) was intraperitoneally injected into mice followed by injection of rat anti-mouse integrin GPIIb/CD41antibody to induce ITP. Peripheral blood was isolated to measure platelet count and spleen mononuclear cells were extracted to measure Th1 and Treg cells along with detecting the levels of IFN-γ, and TGFβ-1 in plasma and CD68 expression in spleen by immuohistochemical staining. Additionally, macrophage cell line RAW264.7 was cultured and treated with DMF followed by analysis of cell apoptosis and cycle, and the expression of FcγRI, FcγRIIb and FcγRIV mRNA.

Results: DMF significantly inhibited antiplatelet antibody-induced platelet destruction, decreased Th1 cells and the expression of T-bet and IFN-γ, upregulated Treg cells and the expression of Foxp3 and TGF-β1 as well as reduced CD68 expression in the spleen of ITP mouse. DMF-treated RAW264.7 cells showed S-phase arrest, increased apoptosis and downregulated expression of FcγRI and FcγRIV. Meanwhile, in vitro treatment of DMF also decreased the expression of cyclin D1 and E2, reduced Bcl-2 level and increased Bax expression and caspase-3 activation.

Conclusions: In conclusion, DMF prevents antibody-mediated platelet destruction in ITP mice possibly through promoting apoptosis, indicating that it might be used as a new approach for the treatment of ITP.
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http://dx.doi.org/10.1186/s12959-021-00314-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8403390PMC
August 2021

Matrine Impairs Platelet Function and Thrombosis and Inhibits ROS Production.

Front Pharmacol 2021 22;12:717725. Epub 2021 Jul 22.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China.

Matrine is a naturally occurring alkaloid and possesses a wide range of pharmacological properties, such as anti-cancer, anti-oxidant, anti-inflammatory effects. However, whether it affects platelet function and thrombosis remains unclear. This study aims to evaluate the effect of matrine on platelet function and thrombus formation. Human platelets were treated with matrine (0-1 mg/ml) for 1 h at 37°C followed by measuring platelet aggregation, granule secretion, receptor expression by flow cytometry, spreading and clot retraction. In addition, matrine (10 mg/kg) was injected intraperitoneally into mice to measure tail bleeding time, arterial and venous thrombus formation. Matrine dose-dependently inhibited platelet aggregation and ATP release in response to either collagen-related peptide (Collagen-related peptide, 0.1 μg/ml) or thrombin (0.04 U/mL) stimulation without altering the expression of P-selectin, glycoprotein Ibα, GPVI, or αIIbβ3. In addition, matrine-treated platelets presented significantly decreased spreading on fibrinogen or collagen and clot retraction along with reduced phosphorylation of c-Src. Moreover, matrine administration significantly impaired the hemostatic function of platelets, arterial and venous thrombus formation. Furthermore, in platelets stimulated with CRP or thrombin, matrine significantly reduced Reactive oxygen species generation, inhibited the phosphorylation level of ERK1/2 (Thr202/Tyr204), p38 (Thr180/Tyr182) and AKT (Thr308/Ser473) as well as increased VASP phosphorylation (Ser239) and intracellular cGMP level. In conclusion, matrine inhibits platelet function, arterial and venous thrombosis, possibly involving inhibition of ROS generation, suggesting that matrine might be used as an antiplatelet agent for treating thrombotic or cardiovascular diseases.
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http://dx.doi.org/10.3389/fphar.2021.717725DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8339414PMC
July 2021

NLRP1 in Bone Marrow Microenvironment Controls Hematopoietic Reconstitution After Transplantation.

Transplant Cell Ther 2021 Jul 21. Epub 2021 Jul 21.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, Jiangsu, China; Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu, China; Key Laboratory of Bone Marrow Stem Cell, Xuzhou, Jiangsu, China. Electronic address:

Pretreatment before transplantation initiates an inflammatory response. Inflammasomes are key regulators of immune and inflammatory responses, but their role in regulating hematopoiesis is unclear. Our study intended to assess the role and mechanism of nucleotide-binding domain and leucine-rich repeat pyrin-domain containing protein 1 (NLRP1) in the bone marrow microenvironment on hematopoiesis regulation. To explore the effects of an absence of NLRP1 on hematopoietic reconstitution, we established a hematopoietic cell transplantation model by infusing bone marrow mononuclear cells of wild-type C57BL/6 mice into either NLRP1 knockout (NLRP1-KO) or wild-type C57BL/6 mice. Using the transplantation model, the role of NLRP1 in the bone marrow microenvironment was determined by flow cytometry, hemacytometry, and hematoxylin and eosin staining. As the major component of the bone marrow microenvironment, mesenchymal stem cells (MSCs) were isolated to analyze the effects of NLRP1 on them by osteogenic and adipogenic induction. Endothelial cells (ECs) were isolated and sorted by magnetic beads. The expression of adhesion molecules and their relationship with nuclear factor kappa B (NF-κB) were measured by immunofluorescence, enzyme-linked immunosorbent assay, and western blot. Finally, the effect of NLRP1-deleted MSCs or ECs on hematopoietic stem and progenitor cells (HSPCs) was examined by establishing co-culture models. Compared with C57BL/6 recipients, reduced inflammatory cell infiltration, decreased levels of proinflammatory cytokines interleukin (IL)-18, IL-1β, IL-6, tumor necrosis factor alpha (TNF-α), and interferon gamma (IFN-γ), together with reduced pathological injury of bone marrow, were observed in NLRP1-KO recipients after transplantation. However, increased HSPC engraftment and hematopoietic reconstitution were detected in NLRP1-KO recipients after transplantation. Furthermore, MSCs isolated from NLRP1-KO mice had decreased osteogenic and adipogenic differentiation and increased proliferation and differentiation of HSPCs. The expression of adhesion molecules in ECs from NLRP1-KO mice was increased due to the promotion of nuclear translocation of NF-κB; these adhesion molecules are critical for hematopoietic stem cell homing. Knockout of NLRP1 in the bone marrow microenvironment could significantly relieve bone marrow inflammatory response and promote hematopoietic reconstitution, perhaps by regulating MSCs and ECs, indicating that NLRP1 might be a target for the treatment of delayed hematopoietic and immune recovery in patients after hematopoietic stem cell transplantation.
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http://dx.doi.org/10.1016/j.jtct.2021.07.016DOI Listing
July 2021

Gamma Radiation Induce Inflammasome Signaling and Pyroptosis in Microvascular Endothelial Cells.

J Inflamm Res 2021 15;14:3277-3288. Epub 2021 Jul 15.

Department of Blood Diseases Institute, Xuzhou Medical University, Xuzhou City, 221002, Jiangsu Province, People's Republic of China.

Introduction: The extend to the clinical benefit of radiation therapy is the inability to eliminate only cancer cells and destroy normal cells such as microvascular endothelial in the vascular niche and turn induced-inflammasome signaling and cell death. These unfortunate injuries generated by ionizing radiation alter the therapeutic window and result in the re-occurrence of the malignancy. Therefore, we engaged in vitro studies by demonstrating radiation-induced inflammasome and cell death in endothelial cells.

Methods: The microvascular endothelial cells were cultured in a sterile dish, then kept in a humidifier of 5% at 37°C for 12 hours/more to attain confluence, and exposed at a dose of 1.8Gy/min achieve the coveted amounts except for the control. The cells were harvested 24 hours post-irradiation.

Results: Our findings indicate that gamma radiation activates the NOD-like receptor (NLR) family of NLRP1 and NLRP3 complex in microvascular endothelial cells. These complexes activate the inactive precursor of caspase-1, which cleaved to bioactive caspase -1 and enhances the production of pro-inflammatory cytokines of interleukin-1β and interleukin-18 that induce the dependent pyroptotic, which results in the production of chemokines, tumor necrosis factor-alpha (TNF-α), and high-mobility group protein-1 (HMGB-1). We also discovered the radiation could directly prompt caspase -1, which auto-cleaved to activate gasdermin D to potentiate pyroptosis independently.

Discussion: Overall, these findings suggested that reducing the unfavorable effect of radiation injuries could be challenging since gamma radiation induces the microvascular endothelial cells to cell death and activates the inflammasome signaling via different pathways.
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http://dx.doi.org/10.2147/JIR.S318812DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8289370PMC
July 2021

Surface site density and utilization of platinum group metal (PGM)-free Fe-NC and FeNi-NC electrocatalysts for the oxygen reduction reaction.

Chem Sci 2020 Oct 13;12(1):384-396. Epub 2020 Oct 13.

The Electrochemical Catalysis, Energy and Materials Science Laboratory, Department of Chemistry, Technische Universität Berlin Straße des 17. 10623 Berlin Germany

Pyrolyzed iron-based platinum group metal (PGM)-free nitrogen-doped single site carbon catalysts (Fe-NC) are possible alternatives to platinum-based carbon catalysts for the oxygen reduction reaction (ORR). Bimetallic PGM-free MM-NC catalysts and their active sites, however, have been poorly studied to date. The present study explores the active accessible sites of mono- and bimetallic Fe-NC and FeNi-NC catalysts. Combining CO cryo chemisorption, X-ray absorption and Fe Mössbauer spectroscopy, we evaluate the number and chemical state of metal sites at the surface of the catalysts along with an estimate of their dispersion and utilization. Fe L-edge X-ray adsorption spectra, Mössbauer spectra and CO desorption all suggested an essentially identical nature of Fe sites in both monometallic Fe-NC and bimetallic FeNi-NC; however, Ni blocks the formation of active sites during the pyrolysis and thus causes a sharp reduction in the accessible metal site density, while with only a minor direct participation as a catalytic site in the final catalyst. We also use the site density utilization factor, , as a measure of the metal site dispersion in PGM-free ORR catalysts. enables a quantitative evaluation and comparison of distinct catalyst synthesis routes in terms of their ratio of accessible metal sites. It gives guidance for further optimization of the accessible site density of M-NC catalysts.
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http://dx.doi.org/10.1039/d0sc03280hDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8179675PMC
October 2020

A novel strategy for isolation of mice bone marrow endothelial cells (BMECs).

Stem Cell Res Ther 2021 05 3;12(1):267. Epub 2021 May 3.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, 221002, China.

Background: In the bone marrow microenvironment (BM), endothelial cells are individual cells that form part of the sinusoidal blood vessels called the "bone marrow endothelial-vascular niche." They account for less than 2% of the bone marrow cells. They play essential functions by generating growth and inhibitory factors that promote the hematopoietic stem cells (HSCs) regulation. In response to inflammatory stimuli, the BMECs increase in proliferation to maintain the blood vessels' integrity within the BM. The inflammatory response releases cytokines such as tumor necrosis factor-alpha (TNF-α) that promote vascular endothelial cells' expansion and upregulation of adhesion molecules (ICAM-1 and VCAM-1, respectively) in the BM. However, the evaluation of mouse BMECs in the bone marrow microenvironment is scared by a lack of mouse bone marrow endothelial cell primary culture METHODS: Two steps approach for isolation of bone marrow endothelial cells (BMECs) from mice. In brief, the bone marrow cells extracted from the mice long bones were cultured overnight with Dulbecco's modified Eagle's medium (DMEM) supplemented with 20% fetal bovine serum (FBS) and antibiotics to separate between marrow-derived adherent and non-adherent cells. The floating cells were discarded, and the adhered section detached with accutase and BMECs selected using CD31 microbeads. The isolated BMECs were cultured in a dish pre-coated with rat-tail collagen type 1 with endothelial cells medium supplement with growth factors. The cells were verified by confocal microscopy for morphology and tube formation by matrigel assay. We validate the cells' purity by flow cytometry, RT-qPCR, immunofluorescence staining, and immunoblotting by established BMEC markers, PECAM-1, VE-cadherin, vascular endothelial cell growth factor receptor-2 (VEGFR2), CD45, E-selectin, and endothelial selectin adhesion molecule (ESAM). Lastly, we characterize BMEC activation with recombinant TNF-α.

Results: Our method clearly defined the cells isolated have the characteristics of BMECs with the expression of CD31, VE-cadherin, E-selectin, VEGFR-2, and ESAM. The cells' response to TNF-α indicates its inflammatory function by increasing proliferation and upregulation of adhesion molecules.

Conclusions: This study outline a simple new technique of isolating mouse BMEC primary culture and a suitable method to evaluate the function and dysregulation of BMEC in in vitro studies using mouse models.
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http://dx.doi.org/10.1186/s13287-021-02352-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8091666PMC
May 2021

PIK3CA Is Regulated by CUX1, Promotes Cell Growth and Metastasis in Bladder Cancer Activating Epithelial-Mesenchymal Transition.

Front Oncol 2020 3;10:536072. Epub 2020 Dec 3.

Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

PIK3CA is a key component of phosphatidylinositol 3-kinase (PI3K) pathway that its involvement in tumorigenesis has been revealed by previous research. However, its functions and potential mechanisms in bladder cancer are still largely undiscovered. Tissue microarray (TMA) with 66 bladder cancer patients was surveyed immunohistochemistry to evaluate the level of PIK3CA and CUX1 and we found upregulation of PIK3CA in bladder cancer tissue and patients with higher level of PIK3CA presented with poorer prognosis. Overly expressed PIK3CA promoted growth, migration, invasion, and metastasis of bladder cancer cells and knockdown of PIK3CA had the opposite effect. Gain-of-function and loss-of-function studies showed that expression was facilitated by CUX1, leading to activation of epithelial-mesenchymal transition (EMT), accompanied by upregulated expression of Snail, β-catenin, Vimentin and downregulated expression of E-cadherin in the bladder cancer cell lines. Besides, over-expressed CUX1 could restore the expression of downregulated Snail, β-catenin, Vimentin and E-cadherin which was induced by PIK3CA knockdown. These results revealed that overexpression in bladder cancer was regulated by the transcription factor CUX1, and exerted its biological effects by activating EMT.
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http://dx.doi.org/10.3389/fonc.2020.536072DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7744743PMC
December 2020

C-C Coupling Reactions for the Synthesis of Two-Dimensional Conjugated Polymers.

Chempluschem 2020 Dec;85(12):2636-2651

Hubei Collaborative Innovation Center for Advanced Organic Chemical Materials, Ministry-of-Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules, Hubei Key Laboratory of Polymer Materials, College of Chemistry and Chemical Engineering, Hubei University, Wuhan, 430062, P. R. China.

Extension of conjugated polymers from 1D to 2D can not only significantly enhance the dissociation of charge and excitons, but also induce other advantages, such as high in-plane mechanical strength, large specific surface area and porosity, and more active centers. 2D conjugated polymers can be divided into C-C bonded 2D polymers based on C-C coupling reactions, and heteroatomic bonded 2D polymers based on reversible heteroatom coupling reactions. C-C bonded 2D polymers are generally more stable than heteroatomic bonded 2D polymers as the latter bonds are easily hydrolyzed. This Review mainly summarizes C-C coupling reactions that are suitable for synthesizing 2D conjugated polymers, and the properties of these 2D conjugated polymers are also introduced.
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http://dx.doi.org/10.1002/cplu.202000643DOI Listing
December 2020

Clodronate-liposomes aggravate irradiation-induced myelosuppression by promoting myeloid differentiation.

Int J Radiat Biol 2021 11;97(2):240-248. Epub 2021 Jan 11.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China.

Purpose: Clodronate-liposomes (Clod-Lip) is an effective candidate drug for treating chronic myelomonocytic leukemia, autoimmune hemolytic anemia and immune thrombocytopenic purpura in mice experiments. But its role in hematopoietic recovery after acute myelosuppression is still unknown. We aim to explore the function and underlining mechanisms of Clod-Lip on hematopoietic reconstitution after sublethal dose irradiation in mice.

Materials And Methods: Mice at 8-10 weeks received a total-body sublethal dose γ-irradiation (TBI) and injected with Clod-Lip or PBS-Liposomes (PBS-Lip) every 4 days after TBI. The survival rate of each group was recorded. Flow cytometry was used to analyze changes in hematopoietic stem cells and their progenies in bone marrow. ELISA and RT-qPCR were used for the analysis of hematopoietic regulatory factors. Regarding IL-1β inhibition, 25 mg/kg diacerein or an equal volume of DMSO was intraperitoneally injected into mice every day after TBI.

Results: In sublethal dose-irradiated mice, Clod-Lip reduced the survival rate, the total number of bone marrow and hematopoietic stem cells, delayed peripheral blood recovery of red blood cells and platelets. However, it could increase the number of CMP, MEP and myeloid cells, which suggested that Clod-Lip could induce HSC to myeloid differentiation in vivo. We further verified that Clod-Lip may induce myeloid differentiation by bone marrow microenvironmental factor IL-1β.

Conclusions: In summary, this study suggested that Clod-Lip may aggravate inhibitor effect of hematopoietic function and promote myeloid differentiation in myelosuppression mice model.
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http://dx.doi.org/10.1080/09553002.2021.1857452DOI Listing
August 2021

High expression of disabled homolog 2-interacting protein contributes to tumor development and proliferation in cutaneous squamous cell carcinoma.

Ann Transl Med 2020 Sep;8(18):1131

Nanhai District People's Hospital, Foshan, China.

Background: Disabled homolog 2-interacting protein (DAB2IP), a Ras GTPase-activating protein, is downregulated in several cancers. Its depletion is involved in tumor cell proliferation, apoptosis, and metastasis, as well as epithelial-mesenchymal transition. The present study aimed to explore the potential role of DAB2IP in cutaneous squamous cell carcinoma (cSCC) and provide a theoretical basis for the diagnosis and targeted therapy of cSCC.

Methods: The clinicopathological features of DAB2IP expression in cSCC were analyzed by immunohistochemistry, and the effects of DAB2IP on SCL-1 cell behavior were determined via genetic interference . SCL-1 cell lines that exhibited reduced expression of DAB2IP and a scrambled shRNA control were constructed using a lentivirus vector-based shRNA technique. RNA extraction, reverse transcription-quantitative PCR (RT-qPCR), MTT assay, colony formation test, cell cycle analysis, apoptosis test, transwell assay, wound-healing assay, invasive assay were used in this study.

Results: The immunohistochemical results demonstrated that the expression of DAB2IP was higher in cSCC tissues than in soft fibroma. The level of DAB2IP expression was associated with the degree of malignancy and the depth of tumor infiltration; however, it had no association with patients' sex, tumor size, location, or phenotype. The results of the MTT, cell cycle, apoptosis, and invasion experiments demonstrated that knockdown of DAB2IP inhibited the viability and invasion of SCL-1 cells .

Conclusions: High expression of DAB2IP may contribute to the development and proliferation of cSCC.
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http://dx.doi.org/10.21037/atm-20-5067DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7576015PMC
September 2020

Real-world investigation of the efficacy and safety of secukinumab for psoriasis treatment in a Chinese population.

Chin Med J (Engl) 2020 Oct 26;134(1):117-119. Epub 2020 Oct 26.

Department of Dermatology, Guangzhou Institute of Dermatology, Guangzhou, Guangdong 510095, China.

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http://dx.doi.org/10.1097/CM9.0000000000001179DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7862816PMC
October 2020

Antibiotics in cultured freshwater products in Eastern China: Occurrence, human health risks, sources, and bioaccumulation potential.

Chemosphere 2021 Feb 28;264(Pt 1):128441. Epub 2020 Sep 28.

College of Marine Ecology and Environment, Shanghai Ocean University, Shanghai, 201306, China.

The adverse effects of antibiotics residues on aquaculture ecosystems and humans raised increasing concerns globally. To assess the occurrence, human health risks, sources, and bioaccumulation potential of antibiotics in cultured freshwater products in Eastern China, 12 and 13 aquaculture ponds were selected in 2018 and 2019, respectively, both covering 8 aquatic species. Concentrations of 12 commonly-used antibiotics were measured in muscle tissue of aquaculture products, water, sediment, and suspended particles. At least two antibiotics were found simultaneously in all muscle tissue samples. The concentrations of most antibiotics in freshwater cultured products were at a medium or lower level in comparison with other studies in China and worldwide, but slightly higher than the concentrations in cultured marine products. The potential risks from the intake of these aquatic products were also evaluated. The results showed limited adverse effects due to the consumption of these products with an exception of fluoroquinolone antibiotics. The bioaccumulation potential from water varied widely in different collection years, but the bioaccumulation factor (BAF) values for antibiotics were all <50 L/kg. BSAF values of antibiotics were all far below 1, except for one site in Zhejiang province in 2018, indicating that the bioavailability from surface sediments was low, in a particular pond environment. The low repeatability of BAF and BSAF calculated in two years indicated a relatively unsteady status in terms of bioaccumulation potential of cultured freshwater ponds yearly.
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http://dx.doi.org/10.1016/j.chemosphere.2020.128441DOI Listing
February 2021

PEDF promotes the repair of bone marrow endothelial cell injury and accelerates hematopoietic reconstruction after bone marrow transplantation.

J Biomed Sci 2020 Sep 1;27(1):91. Epub 2020 Sep 1.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China.

Background: Preconditioning before bone marrow transplantation such as irradiation causes vascular endothelial cells damage and promoting the repair of damaged endothelial cells is beneficial for hematopoietic reconstitution. Pigment epithelium-derived factor (PEDF) regulates vascular permeability. However, PEDF's role in the repair of damaged endothelial cells during preconditioning remains unclear. The purpose of our study is to investigate PEDF's effect on preconditioning-induced damage of endothelial cells and hematopoietic reconstitution.

Methods: Damaged endothelial cells induced by irradiation was co-cultured with hematopoietic stem cells (HSC) in the absence or presence of PEDF followed by analysis of HSC number, cell cycle, colony formation and differentiation. In addition, PEDF was injected into mice model of bone marrow transplantation followed by analysis of bone marrow injury, HSC number and peripheral hematopoietic reconstitution as well as the secretion of cytokines (SCF, TGF-β, IL-6 and TNF-α). Comparisons between two groups were performed by student t-test and multiple groups by one-way or two-way ANOVA.

Results: Damaged endothelial cells reduced HSC expansion and colony formation, induced HSC cell cycle arrest and apoptosis and promoted HSC differentiation as well as decreased PEDF expression. Addition of PEDF increased CD144 expression in damaged endothelial cells and inhibited the increase of endothelial permeability, which were abolished after addition of PEDF receptor inhibitor Atglistatin. Additionally, PEDF ameliorated the inhibitory effect of damaged endothelial cells on HSC expansion in vitro. Finally, PEDF accelerated hematopoietic reconstitution after bone marrow transplantation in mice and promoted the secretion of SCF, TGF-β and IL-6.

Conclusions: PEDF inhibits the increased endothelial permeability induced by irradiation and reverse the inhibitory effect of injured endothelial cells on hematopoietic stem cells and promote hematopoietic reconstruction.
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http://dx.doi.org/10.1186/s12929-020-00685-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466818PMC
September 2020

Tacrolimus ameliorates thrombocytopenia in an ITP mouse model.

Ann Hematol 2020 Oct 29;99(10):2315-2322. Epub 2020 Jul 29.

Blood Diseases Institute, Xuzhou Medical University, 84 West Huaihai Rd, Quanshan District, Xuzhou, 221002, Jiangsu, China.

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by lower platelet count resulting from immune cells-mediated platelet clearance. Tacrolimus is an immunosuppressive agent which selectively inhibits T cell activation. Whether tacrolimus plays a role in ITP remains unclear. This study aimed to investigate the effect of tacrolimus on ITP in mice. An ITP mouse model was established by injection of rat anti-mouse integrin GPIIb/CD41 immunoglobulin and treated with tacrolimus followed by isolation of peripheral blood mononuclear cells and plasma. The mRNA expression of T-bet, GATA3, and Foxp3 was measured by RT-PCR, and level of IFN-γ, IL-12p70, IL-4, IL-13, and TGF-β in plasma was measured by ELISA. Tacrolimus inhibited antiplatelet antibody-mediated platelet clearance in ITP mouse model. Meanwhile, tacrolimus-treated ITP mice displayed a significant decrease in the mRNA expression of T-bet and plasma level of IFN-γ and IL-12p70 compared with ITP mice but without differences when compared with normal mice. Furthermore, the expression of GATA3, Foxp3, and plasma level of IL-4 and TGF-β were upregulated in tacrolimus-treated ITP mice without significant differences to normal mice (except TGF-β). Tacrolimus prevents antiplatelet antibody-mediated thrombocytopenia in ITP mice possibly through regulating T cell differentiations, suggesting it might be a novel approach for preventing ITP.
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http://dx.doi.org/10.1007/s00277-020-04203-2DOI Listing
October 2020

P-block single-metal-site tin/nitrogen-doped carbon fuel cell cathode catalyst for oxygen reduction reaction.

Nat Mater 2020 Nov 13;19(11):1215-1223. Epub 2020 Jul 13.

Department of Chemistry, The Electrochemical Energy, Catalysis and Material Science Laboratory, Chemical Engineering Division, Technical University Berlin, Berlin, Germany.

This contribution reports the discovery and analysis of a p-block Sn-based catalyst for the electroreduction of molecular oxygen in acidic conditions at fuel cell cathodes; the catalyst is free of platinum-group metals and contains single-metal-atom actives sites coordinated by nitrogen. The prepared SnNC catalysts meet and exceed state-of-the-art FeNC catalysts in terms of intrinsic catalytic turn-over frequency and hydrogen-air fuel cell power density. The SnNC-NH catalysts displayed a 40-50% higher current density than FeNC-NH at cell voltages below 0.7 V. Additional benefits include a highly favourable selectivity for the four-electron reduction pathway and a Fenton-inactive character of Sn. A range of analytical techniques combined with density functional theory calculations indicate that stannic Sn(IV)N single-metal sites with moderate oxygen chemisorption properties and low pyridinic N coordination numbers act as catalytically active moieties. The superior proton-exchange membrane fuel cell performance of SnNC cathode catalysts under realistic, hydrogen-air fuel cell conditions, particularly after NH activation treatment, makes them a promising alternative to today's state-of-the-art Fe-based catalysts.
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http://dx.doi.org/10.1038/s41563-020-0717-5DOI Listing
November 2020

Comparison of survival outcomes of locally advanced cervical cancer by histopathological types in the surveillance, epidemiology, and end results (SEER) database: a propensity score matching study.

Infect Agent Cancer 2020 13;15:33. Epub 2020 May 13.

1Department of Radiation Therapy, The Third Affiliated Hospital of Kunming Medical University, Kunming, 650118 Yunnan China.

Background: There has been limited research on the comparison of squamous cell carcinoma (SCC) and adenocarcinoma (AC) of cervical cancer and that lack of information may have significant bearing on the treatment of patients. We compared survival outcomes between squamous cell carcinoma and adenocarcinoma in locally advanced cervical cancer patients and examined factors related to the prognosis of cervical cancer.

Methods: We identified 4131 patients with stage IB2-IVA cervical cancer patients diagnosed between 2010 and 2015 by using the Surveillance, Epidemiology, and End Results (SEER) database. Variables related to the prognosis of cervical cancer were compared using both univariate and multivariate Cox models and log-rank method before and after propensity score matching. We compared the efficacy of radiotherapy alone to radiotherapy combined with chemotherapy or/and surgery in overall survival of SCC and AC.

Results: Our sample included 3385 patients with SCC (81.9%) and 746 patients with AC (18.1%). The 5-year overall survival on comparing the squamous cell carcinoma group and adenocarcinoma group was not significant ( > 0.05). Using propensity score matching, 676 pairs of patients were selected. The 5-year overall survival of matched patients did not differ significantly ( > 0.05). Histology was not independently associated with overall survival in multivariate Cox model (P > 0.05). Factors affecting overall survival included FIGO stage IVA ( < 0.05), chemotherapy (P < 0.05), and external radiation combined with brachytherapy (P < 0.05). Patients with SCC that were treated with radiation alone had significantly worse OS than AC patients receiving radiation only ( < 0.05).

Conclusions: The OS in AC of the cervix is similar to that SCC in when treated with radiotherapy combined with chemotherapy and/or surgery but better when treated with radiation alone.
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http://dx.doi.org/10.1186/s13027-020-00299-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7222537PMC
May 2020

p47phox deficiency impairs platelet function and protects mice against arterial and venous thrombosis.

Redox Biol 2020 07 11;34:101569. Epub 2020 May 11.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China; Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, China. Electronic address:

NADPH oxidase-derived reactive oxygen species (ROS) regulates platelet function and thrombosis. It remains controversial regarding NOX2's role in platelet function. As a regulatory subunit for NOX2, whether p47phox regulates platelet function remains unclear. Our study intends to evaluate p47phox's role in platelet function. Platelets were isolated from wild-type or p47phox mice followed by analysis of platelet aggregation, granule secretion, surface receptors expression, spreading, clot retraction and ROS generation. Additionally, in vivo hemostasis, arterial and venous thrombosis was assessed. Moreover, human platelets were treated with PR-39 to inhibit p47phox activity followed by analysis of platelet function. p47phox deficiency significantly prolonged tail-bleeding time, delayed arterial and venous thrombus formation in vivo as well as reduced platelet aggregation, ATP release and αIIbβ3 activation. In addition, p47phox platelets presented impaired spreading on fibrinogen or collagen and defective clot retraction concomitant with decreased phosphorylation of Syk and PLCγ2. Moreover, CRP or thrombin-stimulated p47phox platelets displayed reduced intracellular ROS generation which was further decreased after inhibition of NOX1. Meanwhile, p47phox deficiency increased VASP phosphorylation and decreased phosphorylation of ERK1/2, p38, ERK5 and JNK without affecting AKT and c-PLA2 phosphorylation. Furthermore, p47phox translocates to membrane to interact with both NOX1 and NOX2 after stimulation with CRP or thrombin. Finally, inhibition of p47phox activity by PR-39 reduced ROS generation, platelet aggregation and clot retraction in human platelets. In conclusion, p47phox regulates platelet function, arterial and venous thrombus formation and ROS generation, indicating that p47phox might be a novel therapeutic target for treating thrombotic or cardiovascular diseases.
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http://dx.doi.org/10.1016/j.redox.2020.101569DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7231845PMC
July 2020

Salidroside inhibits platelet function and thrombus formation through AKT/GSK3β signaling pathway.

Aging (Albany NY) 2020 04 30;12(9):8151-8166. Epub 2020 Apr 30.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China.

Salidroside is the main bioactive component in and possesses multiple biological and pharmacological properties. However, whether salidroside affects platelet function remains unclear. Our study aims to investigate salidroside's effect on platelet function. Human or mouse platelets were treated with salidroside (0-20 μM) for 1 hour at 37°C. Platelet aggregation, granule secretion, and receptors expression were measured together with detection of platelet spreading and clot retraction. In addition, salidroside (20 mg/kg) was intraperitoneally injected into mice followed by measuring tail bleeding time, arterial and venous thrombosis. Salidroside inhibited thrombin- or CRP-induced platelet aggregation and ATP release and did not affect the expression of P-selectin, glycoprotein (GP) Ibα, GPVI and αβ. Salidroside-treated platelets presented decreased spreading on fibrinogen or collagen and reduced clot retraction with decreased phosphorylation of c-Src, Syk and PLCγ2. Additionally, salidroside significantly impaired hemostasis, arterial and venous thrombus formation in mice. Moreover, in thrombin-stimulated platelets, salidroside inhibited phosphorylation of AKT (T308/S473) and GSK3β (Ser9). Further, addition of GSK3β inhibitor reversed the inhibitory effect of salidroside on platelet aggregation and clot retraction. In conclusion, salidroside inhibits platelet function and thrombosis via AKT/GSK3β signaling, suggesting that salidroside may be a novel therapeutic drug for treating thrombotic or cardiovascular diseases.
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http://dx.doi.org/10.18632/aging.103131DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7244060PMC
April 2020

Reference gene selection and validation for mRNA expression analysis by RT-qPCR in murine M1- and M2-polarized macrophage.

Mol Biol Rep 2020 Apr 19;47(4):2735-2748. Epub 2020 Mar 19.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, 221002, China.

Murine bone marrow-derived macrophages (M0) and M1- and M2-polarized macrophages are being widely used as a laboratory model for polarized macrophages related molecular mechanism analysis. Gene expression analysis based on reference gene normalization using RT-qPCR was a powerful way to explore the molecular mechanism. But little is known about reference genes in these cell models. So, the goal of this study was to identify reference genes in these types of macrophages. Candidate reference genes in murine bone marrow-derived and polarized macrophages were selected from microarray data using Limma linear model method and evaluated by determining the stability value using five algorithms: BestKeeper, NormFinder, GeNorm, Delta CT method, and RefFinder. Finally, the selected stable reference genes were validated by testing three important immune and inflammatory genes (NLRP1, IL-1β, and TNF-α) in the cell lines. Our study has clearly shown that Ubc followed by Eef1a1 and B2m respectively were recognized as the three ideal reference genes for gene expression analysis in murine bone marrow-derived and polarized macrophages. When three reference genes with strong different stability were used for validation, a large variation of a gene expression level of IL-1β, TNF-α and NLRP1 were obtained which provides clear evidence of the need for careful selection of reference genes for RT-qPCR analysis. Normalization of mRNA expression level with Ubc rather than Actb or Gusb by qPCR in macrophages and polarized macrophages is required to ensure the accuracy of the qPCR analysis.
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http://dx.doi.org/10.1007/s11033-020-05372-zDOI Listing
April 2020

Thalidomide prevents antibody-mediated immune thrombocytopenia in mice.

Thromb Res 2019 Nov 22;183:69-75. Epub 2019 Oct 22.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou 221002, China; Department of Hematology, the Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou 221002, China. Electronic address:

Immune thrombocytopenia (ITP) is a heterogeneous autoimmune disorder characterized by immune-mediated platelet destruction, leading to lower platelet count. Thalidomide is considered as a novel immunomodulatory drug for treating several autoimmune diseases. Whether thalidomide can ameliorate ITP remains unclear. This study aims to evaluate the effect of thalidomide on ITP mouse model. ITP mouse model was established through intraperitoneal injection of rat anti-mouse integrin GPIIb/CD41 immunoglobulin. Thalidomide (10, 20 or 50 mg/kg body weight) was intraperitoneally injected into mice followed by antibody injection. Then, peripheral blood and plasma was isolated for analysis of platelet count and the level of IFN-γ and IL-17 in plasma. Meanwhile, spleen was extracted to measure the expression of CD68, a macrophage marker. In addition, macrophage cell line RAW264.7 was cultured and treated with thalidomide followed by analysis of cell viability, apoptosis as well as cell cycle. Thalidomide prevented antiplatelet antibody-mediated platelet destruction in ITP mouse model. Compared with vehicle (phosphate-buffered saline), thalidomide significantly inhibited the secretion of IFN-γ and IL-17 in ITP mouse and reduced the expression of CD68 in spleen. After thalidomide treatment, the cell viability of RAW264.7 cell was significantly reduced and the cell number in S phase was also significantly decreased. In addition, the expression of cyclin E2 was significantly reduced. In conclusion, thalidomide prevents antiplatelet antibody-mediated platelet destruction in ITP mouse possibly through reducing the number of macrophages, suggesting that it might be a novel approach for treating ITP.
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http://dx.doi.org/10.1016/j.thromres.2019.09.035DOI Listing
November 2019

Mechanistic reaction pathways of enhanced ethylene yields during electroreduction of CO-CO co-feeds on Cu and Cu-tandem electrocatalysts.

Nat Nanotechnol 2019 11 7;14(11):1063-1070. Epub 2019 Oct 7.

Department of Chemistry, Chemical Engineering Division, Technical University Berlin, Berlin, Germany.

Unlike energy efficiency and selectivity challenges, the kinetic effects of impure or intentionally mixed CO feeds on the catalytic reactivity of the direct electrochemical CO reduction reaction (CO2RR) have been poorly studied. Given that industrial CO feeds are often contaminated with CO, a closer investigation of the CO2RR under CO/CO co-feed conditions is warranted. Here, we report mechanistic insights into the CO2RR reactivity of CO/CO co-feeds on Cu-based nanocatalysts. Kinetic isotope-labelling experiments-performed in an operando differential electrochemical mass spectrometry capillary flow cell with millisecond time resolution-showed an unexpected enhanced production of CH, with a yield increase of almost 50%, from a cross-coupled CO-CO reactive pathway. The results suggest the absence of site competition between CO and CO molecules on the reactive surface at the reactant-specific sites. The practical significance of sustained local interfacial CO partial pressures under CO depletion is demonstrated by metallic/non-metallic Cu/Ni-N-doped carbon tandem catalysts. Our findings show the mechanistic origin of improved C product formation under co-feeding, but also highlight technological opportunities of impure CO/CO process feeds for HO/CO co-electrolysers.
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http://dx.doi.org/10.1038/s41565-019-0551-6DOI Listing
November 2019

All-Trans Retinoic Acid Impairs Platelet Function and Thrombus Formation and Inhibits Protein Kinase CßI/δ Phosphorylation.

Thromb Haemost 2019 10 1;119(10):1655-1664. Epub 2019 Aug 1.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China.

All-trans retinoic acid (ATRA) is widely used for induction of complete remission in patients with acute promyelocytic leukemia (APL). ATRA also regulates protein kinase C (PKC) activity. Therapeutic use of ATRA reportedly interferes with hemostatic function in APL patients, including effects on coagulation or other vascular cells, although effects of ATRA on platelets remain unclear. This study aims to investigate the effect of therapeutic-relevant doses of ATRA on platelet function. Human platelets were preincubated with ATRA (0-20 μM) for 1 hour at 37°C, followed by analysis of aggregation, granule secretion, receptor expression by flow cytometry, platelet spreading, or clot retraction. Additionally, ATRA (10 mg/kg) was injected intraperitoneally into mice and tail bleeding time and arterial thrombus formation were evaluated. ATRA inhibited platelet aggregation and adenosine triphosphate release induced by collagen (5 μg/mL) or thrombin (0.05 U/mL) in a dose-dependent manner without affecting P-selectin expression or surface levels of glycoprotein (GP) Ibα, GPVI, or αβ. ATRA-treated platelets demonstrated reduced spreading on immobilized fibrinogen or collagen and reduced thrombin-induced clot retraction together with reduced phosphorylation of Syk and PLCγ2. In addition, ATRA-treated mice displayed significantly impaired hemostasis and arterial thrombus formation in vivo. Further, in platelets stimulated with either collagen-related peptide or thrombin, ATRA selectively inhibited phosphorylation of PKCßI (Ser661) and PKCδ (Thr505), but not PKCα or PKCßII phosphorylation (Thr638/641). In conclusion, ATRA inhibits platelet function and thrombus formation, possibly involving direct or indirect inhibition of PKCßI/δ, indicating that ATRA might be beneficial for the treatment of thrombotic or cardiovascular diseases.
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http://dx.doi.org/10.1055/s-0039-1693737DOI Listing
October 2019

Activity-Selectivity Trends in the Electrochemical Production of Hydrogen Peroxide over Single-Site Metal-Nitrogen-Carbon Catalysts.

J Am Chem Soc 2019 Aug 29;141(31):12372-12381. Epub 2019 Jul 29.

Department of Chemistry , Technical University of Berlin , 10623 Berlin , Germany.

Nitrogen-doped carbon materials featuring atomically dispersed metal cations (M-N-C) are an emerging family of materials with potential applications for electrocatalysis. The electrocatalytic activity of M-N-C materials toward four-electron oxygen reduction reaction (ORR) to HO is a mainstream line of research for replacing platinum-group-metal-based catalysts at the cathode of fuel cells. However, fundamental and practical aspects of their electrocatalytic activity toward two-electron ORR to HO, a future green "dream" process for chemical industry, remain poorly understood. Here we combined computational and experimental efforts to uncover the trends in electrochemical HO production over a series of M-N-C materials (M = Mn, Fe, Co, Ni, and Cu) exclusively comprising atomically dispersed M-N sites from molecular first-principles to bench-scale electrolyzers operating at industrial current density. We investigated the effect of the nature of a 3d metal within a series of M-N-C catalysts on the electrocatalytic activity/selectivity for ORR (HO and HO products) and HO reduction reaction (HORR). Co-N-C catalyst was uncovered with outstanding HO productivity considering its high ORR activity, highest HO selectivity, and lowest HORR activity. The activity-selectivity trend over M-N-C materials was further analyzed by density functional theory, providing molecular-scale understandings of experimental volcano trends for four- and two-electron ORR. The predicted binding energy of HO* intermediate over Co-N-C catalyst is located near the top of the volcano accounting for favorable two-electron ORR. The industrial HO productivity over Co-N-C catalyst was demonstrated in a microflow cell, exhibiting an unprecedented production rate of more than 4 mol peroxide g h at a current density of 50 mA cm.
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http://dx.doi.org/10.1021/jacs.9b05576DOI Listing
August 2019

NLRP6 deficiency aggravates liver injury after allogeneic hematopoietic stem cell transplantation.

Int Immunopharmacol 2019 Sep 10;74:105740. Epub 2019 Jul 10.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou 221002, China; Department of Hematology, The Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, China; School of Medical Technology, Xuzhou Medical University, 221004, China. Electronic address:

This study aims to observe the expression and role of NLRP6 in liver injury after allogeneic hematopoietic stem cell transplantation (Allo-HSCT). Allo-HSCT model was established through infusion of 5 × 10 bone marrow mononuclear cells into whole body irradiated mice. On days 7, 14, 21 and 28 after transplantation, the peripheral blood was collected to detect liver function. The liver of the mice was obtained to assess the pathological changes of liver tissues after allo-HSCT by H&E staining and Mason staining. Meanwhile, expression of NLRP6, phosphorylated p38-MAPK and IκBα, caspase-1 and NLRP3 in liver were detected by Western blot. ELISA was used for detection of the level of interleukin (IL)-1β, IL-18, tumor necrosis factor (TNF)-α, IL-6, myeloperoxidase (MPO) and tumor growth factor (TGF)-β1. Increased expression of NLRP6, phosphorylated Iκbα, phosphorylated p38-MAPK, pro-caspase-1, and p20, in liver tissue with injury and fibrosis in mice after allo-HSCT were observed. Meanwhile, the level of IL-1β, IL-18, IL-6 and TNF-α was also increased. However, NLRP6 mice showed more severe liver damage and liver fibrosis after transplantation together with higher level of phosphorylated Iκbα, phosphorylated p38-MAPK, Pro-caspase-1, p20 expression as well as IL-1β, IL-18, IL-6, and TNF-α secretion compared with wide-type. Interestingly, the expression of NLRP3 in the liver of NLRP6 mice was significantly higher than that of wild-type. In conclusion, the expression of NLRP6 in host's liver is associated with liver injury after allo-HSCT. NLRP6 deficiency in host's liver leads to more severe liver damage, indicating a protective role of NLRP6 in host's liver to liver damage after allo-HSCT.
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http://dx.doi.org/10.1016/j.intimp.2019.105740DOI Listing
September 2019

LncRNA GASL1 inhibits growth and promotes expression of apoptosis-associated proteins in prostate carcinoma cells through GLUT-1.

Oncol Lett 2019 Jun 12;17(6):5327-5334. Epub 2019 Apr 12.

Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China.

Growth-arrest-associated long non-coding (lnc)RNA 1 (GASL1) is as newly identified lncRNA that is associated with liver cancer. The present study aimed to investigate the role of GASL1 in prostate carcinoma (PC). Expression levels of GASL1 in prostate tissues and sera from patients with PC and from healthy subjects were detected by reverse transcription-quantitative polymerase chain reaction. Receiver operating characteristic and survival curve analyses were performed to evaluate the diagnostic and prognostic values of GASL1 for patients with PC. A GASL1 expression vector was transfected into PC cells prior to assessment of cell proliferation and expression of B cell lymphoma 2 (Bcl-2) and glucose transporter 1 (GLUT-1) by Cell Counting Kit-8 and western blotting, respectively. The results demonstrated that GASL1 was significantly downregulated in the tissue and serum of patients with PC compared to those of healthy subjects. In addition, GASL1 was used to distinguish patients with PC from healthy controls, and low expression levels of GASL1 were associated with short survival time. Expression levels of GASL1 were significantly associated with tumor size. GASL1 overexpression inhibited PC cell growth. Overexpression of GASL1 upregulated Bcl-2 expression and downregulated GLUT-1 expression. In conclusion, these data suggested that lncRNA GASL1 may inhibit PC cell proliferation by targeting GLUT-1.
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http://dx.doi.org/10.3892/ol.2019.10244DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6507392PMC
June 2019

MicroRNA‑122 downregulates Rho‑associated protein kinase 2 expression and inhibits the proliferation of prostate carcinoma cells.

Mol Med Rep 2019 May 27;19(5):3882-3888. Epub 2019 Feb 27.

Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China.

MicroRNA‑122 (miR‑122) has been reported to be involved in the pathogenesis of several types of malignancies; however, its role in prostate carcinoma remains unknown. Thus, the current study aimed to investigate the functionality of miR‑122 in prostate carcinoma. Clinical data of 54 patients with prostate carcinoma who were diagnosed and treated in Union Hospital (Wuhan, China) between January 2011 and January 2013 were retrospectively analyzed. The expression levels of miR‑122 and Rho‑associated protein kinase 2 (ROCK2) in prostate tumor and adjacent healthy tissues of patients, as well as in the serum of prostate carcinoma patients and healthy controls, were detected by reverse transcription‑quantitative polymerase chain reaction. Receiver operating characteristic curve and survival curve analyses were used to examine the diagnostic and prognostic values of serum miR‑122 for prostate carcinoma. In addition, miR‑122 mimic was transfected into prostate carcinoma cells, and the effects on cell proliferation and ROCK2 expression were explored by Cell Counting Kit‑8 and western blot assays, respectively. It was observed that miR‑122 was downregulated and ROCK2 was upregulated in tumor tissues as compared with their levels in adjacent healthy tissues. miR‑122 level in the serum was also markedly lower in prostate carcinoma patients in comparison with that in healthy controls. Furthermore, a low serum level of miR‑122 was found to effectively distinguish the prostate carcinoma patients from healthy controls and to be an indicator of poor survival. In prostate carcinoma cells, miR‑122 overexpression inhibited the proliferation and the expression of ROCK2. Taken together, miR‑122 may inhibit the proliferation of prostate carcinoma cells possibly by downregulating ROCK2 expression.
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http://dx.doi.org/10.3892/mmr.2019.9995DOI Listing
May 2019

Genome-wide identification, phylogeny, and expression analysis of Sec14-like PITP gene family in sugarcane.

Plant Cell Rep 2019 May 12;38(5):637-655. Epub 2019 Feb 12.

Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.

Key Message: Six Sec14-like PITP genes from sugarcane were identified, two of them were cloned, and their biological functions were characterized indicating their involvement in plant defense against biotic and abiotic stresses. Sec14, a phosphatidylinositol transfer protein (PITP) is widely present in eukaryotes. In this study, the structure and expression patterns of six Sec14-like PITP genes (ScSEC14-1, ScSEC14p, ScSFH1, ScSFH2, ScPATL1, and ScPATL2) from sugarcane were analyzed, and two of them (ScSEC14-1 and ScSEC14p) were cloned and functionally verified. Phylogenetic analysis divided these genes into four groups, including group I (ScSFH1 and ScSFH2), group II (ScPATL1 and ScPATL2), Group III (ScSEC14p), and group V (ScSEC14-1). qRT-PCR analysis showed tissue-specific expression of these genes, primarily in the root, leaf, and bud tissues. They responded differently to SA, MeJA, and ABA stresses. ScSEC14-1, ScSEC14p, and ScSFH2 were upregulated by CuCl and CdCl, while ScSEC14-1, ScSFH1, ScSFH2, and ScPATL1 were upregulated by PEG and NaCl. When infected by Sporisorium scitamineum, the transcripts of ScSFH1, ScSFH2, ScPATL1, and ScPATL2 were upregulated in the resistant genotype Yacheng 05-179, while those of ScSEC14-1 and ScSEC14p were upregulated in the susceptible genotype ROC22. Subcellular localization showed that ScSEC14-1 and ScSEC14p were mainly localized in the plasma membrane and cytoplasm. Enhanced growth of Escherichia coli BL21 cells expressing ScSEC14-1 and ScSEC14p showed high tolerance to NaCl and mannitol stresses. The transient overexpression of ScSEC14-1 and ScSEC14p in Nicotiana benthamiana leaves enhanced its resistance to the infection of tobacco pathogens Ralstonia solanacearum and Fusarium solani var. coeruleum. We can conclude the involvement of ScSEC14-1 and ScSEC14p in the defense against biotic and abiotic stresses, which should facilitate further research on Sec14-like PITP gene family, especially its regulatory mechanisms in sugarcane.
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http://dx.doi.org/10.1007/s00299-019-02394-1DOI Listing
May 2019

Construction of immune-related risk signature for renal papillary cell carcinoma.

Cancer Med 2019 01 5;8(1):289-304. Epub 2018 Dec 5.

Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.

The kidney renal papillary cell carcinoma (KIRP) is a relatively rare type of kidney cancer. There has been no investigation to find a robust signature to predict the survival outcome of KIRP patients in the aspect of tumor immunology. In this study, 285 KIRP samples from The Cancer Genome Atlas (TCGA) were randomly divided into training and testing set. A total of 1534 immune-related genes from The Immunology Database and Analysis Portal (ImmPort) were used as candidates to construct the signature. Using univariate Cox analysis, we evaluated the relationship between overall survival and immune-related genes expression and found 272 immune-related genes with predicting prognostic ability. In order to construct an efficient predictive model, the Cox proportional hazards model with an elastic-net penalty was used and identified 23 groups after 1000 iterations. As a result, 15-genes model showing more stable than other gene groups was chosen to construct our immune-related risk signature. In line with our expectations, the signature can independently predict the survival outcome of KIRP patients. Patients with high-immune risk were found correlated with advanced stage. We also found that the high-immune risk patients with higher PBRM1 and SETD2 mutations, increasing chromosomal instability, together with the gene set enrichment analysis (GSEA) results showing intensive connection of our signature with immune pathways. In conclusion, our study constructs a robust 15-gene signature for predicting KIRP patients' survival outcome on the basis of tumor immune environment and may provide possible relationship between prognosis and immune-related biological function.
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http://dx.doi.org/10.1002/cam4.1905DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6346237PMC
January 2019

Increased GPIbα shedding from platelets treated with immune thrombocytopenia plasma.

Int Immunopharmacol 2019 Jan 13;66:91-98. Epub 2018 Nov 13.

Blood Diseases Institute, Xuzhou Medical University, Xuzhou, China; Department of Hematology, the Affiliated Hospital of Xuzhou Medical University, Xuzhou, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, China. Electronic address:

Immune thrombocytopenia (ITP) is a heterogeneous autoimmune disease, characterized by accelerated platelet destruction/clearance or decreased platelet production. ADAM17-mediated platelet receptor GPIbα extracellular domain shedding has been shown to be involved in platelet clearance. Whether GPIbα shedding participates in the pathogenesis of ITP remains poorly understood. This study aims to investigate the role of GPIbα shedding in the development of ITP via incubating normal platelets with ITP plasma to mimic ITP in vivo environment. Plasma was isolated from ITP patients or healthy control and incubated with platelets in vitro followed by measuring GPIbα expression by flow cytometry and western blot, ADAM17 expression by western blot, ROS generation and platelet activation by flow cytometry. Compared with control plasma, ITP plasma-treated platelet displayed significantly reduced GPIbα surface expression, increased ADAM17 expression and ROS generation. However, metalloproteinase inhibitor GM6001 blocked the ITP-plasma-induced decrease in GPIbα surface expression, increase in ADAM17 expression and platelet activation. In addition, inhibitors of NADPH oxidase or mitochondria respiration significantly inhibited ROS generation from ITP plasma-treated platelets. Moreover, ROS inhibition or blocking FcγRIIa attenuated the decrease in GPIbα surface expression, platelet activation and ROS generation (for blocking FcγRIIa) in ITP plasma-treated platelets. In conclusion, ITP plasma induces platelet receptor GPIbα extracellular domain shedding, suggesting that it might participate in the pathogenesis of ITP and targeting it might be a novel approach for treating ITP.
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http://dx.doi.org/10.1016/j.intimp.2018.11.011DOI Listing
January 2019

Platycodin D inhibits platelet function and thrombus formation through inducing internalization of platelet glycoprotein receptors.

J Transl Med 2018 11 15;16(1):311. Epub 2018 Nov 15.

Blood Diseases Institute, Xuzhou Medical University, 84 West Huaihai Road, Xuzhou, 221002, China.

Background: Platycodin D (PD) is one of the major bioactive components of the roots of Platycodon grandiflorum and possesses multiple biological and pharmacological properties, such as antiviral, anti-inflammatory, and anti-cancer activities. However, whether it affects platelet function remains unclear. This study aims to evaluate the role of PD in platelet function and thrombus formation.

Methods: Platelets were treated with PD followed by measuring platelet aggregation, activation, spreading, clot retraction, expression of glycoprotein receptors. Moreover, mice platelets were treated with PD and infused into wild-type mice for analysis of in vivo hemostasis and arterial thrombosis.

Results: Platycodin D treatment significantly inhibited platelet aggregation in response to collagen, ADP, arachidonic acid and epinephrine, reduced platelet P-selectin expression, integrin αβ activation, spreading on fibrinogen as well as clot retraction, accompanied with decreased phosphorylation of Syk and PLCγ2 in collagen-related peptide or thrombin-stimulated platelets. Moreover, PD-treated mice platelets presented significantly impaired in vivo hemostasis and arterial thrombus formation. Interestingly, PD induced internalization of glycoprotein receptors αβ, GPIbα and GPVI. However, GM6001, cytochalasin D, BAPTA-AM and wortmannin did not prevent PD-induced internalization of receptors.

Conclusions: Our study demonstrates that PD inhibits platelet aggregation, activation and impairs hemostasis and arterial thrombosis, suggesting it might be a potent anti-thrombotic drug.
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http://dx.doi.org/10.1186/s12967-018-1688-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6238268PMC
November 2018
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