Adhesion between EVs and tumor cells facilitated EV-encapsulated doxorubicin delivery via ICAM1.

Doxorubicin (Dox) is an anti-tumor drug with a broad spectrum, whereas the cardiotoxicity limits its further application. In clinical settings, liposome delivery vehicles are used to reduce Dox cardiotoxicity. Here, we substitute extracellular vesicles (EVs) for liposomes and deeply investigate the mechanism for EV-encapsulated Dox delivery. The results demonstrate that EVs dramatically increase import efficiency and anti-tumor effects of Dox in vitro and in vivo, and the efficiency increase benefits from its unique entry pattern. Dox-loading EVs repeat a "kiss-and-run" motion before EVs internalization. Once EVs touch the cell membrane, Dox disassociates from EVs and directly enters the cytoplasm, leading to higher and faster Dox import than single Dox. This unique entry pattern makes the adhesion between EVs and cell membrane rather than the total amount of EV internalization the key factor for regulating the Dox import. Furthermore, we recognize ICAM1 as the molecule mediating the adhesion between EVs and cell membranes. Interestingly, EV-encapsulated Dox can induce ICAM1 expression by irritating IFN-γ and TNF-α secretion in TME, thereby increasing tumor targeting of Dox-loading EVs. Altogether, EVs and EV-encapsulated Dox synergize via ICAM1, which collectively enhances the curative effects for tumor treatment.

Efficient Hydrogen Production over Molybdenum Tungsten Bimetallic Oxide NF/PMonW12-n Catalyst on Nickel Foam.

Developing inexpensive, efficient, and stable catalysts is crucial for reducing the cost of electrolytic hydrogen production. Recently, polyoxometalates (POMs) have gained attention and widespread use due to their excellent electrocatalytic properties. This study designed and synthesized three composite materials, NF/PMonW12-n, by using phosphomolybdic-tungstic heteropolyacids as precursors to grow in situ on nickel foam via the hydrothermal process and subsequent calcination. Then, their catalytic performances are systematically investigated. This work demonstrates that the NF/PMonW12-n catalysts generate more low valent oxides under the synergistic effect of Mo and W, further enhancing activity for hydrogen evolution reaction (HER). Among these electrocatalysts, NF/PMo6W6 exhibits the perfect HER performance, η10 is only 74 mV. It also shows great stability during long-term electrolysis. The current study introduces a fresh approach for producing electrocatalysts that are both cost-effective and highly efficient.

Ginsenoside Rb1 enhanced immunity and altered the gut microflora in mice immunized by H1N1 influenza vaccine.

Background: Influenza is an acute infectious respiratory disease caused by the influenza virus that seriously damages human health, and the essential way to prevent influenza is the influenza vaccine. Vaccines without adjuvants produce insufficient specific antibodies and therefore require adjuvants to boost antibody titers. Microbes and hosts are a community that needs to "promote bacteria," which could provide new value for the immune effect. Methods: (1) The H1N1 influenza vaccine, in combination with Ginsenoside Rb1, was co-injected into mice intraperitoneally (I.P.). Then, immunoglobulin G and antibody subtype levels were tested by enzyme-linked immunosorbent assay (ELISA). Moreover, mice were infected with a lethal dose of the H1N1 influenza virus (A/Michigan/45/2015), and survival status was recorded for 14 days. Lung tissues were stained by hematoxylin and eosin (H&E), and ELISA detected inflammatory factor expression levels. (2) Mice were immunized with Ginsenoside Rb1 combined with quadrivalent influenza inactivated vaccine(IIV4), and then IgG levels were measured by ELISA. (3) Fresh stool was collected for fecal 16S rDNA analysis. Results: Ginsenoside Rb1 boosted IgG and antibody subtypes in the H1N1 influenza vaccine, improved survival of mice after virus challenge, attenuated lung histopathological damage, and reduced inflammatory cytokines expression in IL-6 and TNF-α. The results of 16S rDNA showed that Rb1 decreased species diversity but increased species richness compared to the PBS group and increased the abundance of Akkermansiaceae and Murbaculaceae at the Family and Genus levels compared with the HA+Alum group. Conclusion: Ginsenoside Rb1 has a boosting effect on the immune efficacy of the H1N1 influenza vaccine and is promising as a novel adjuvant to regulate the microecological balance and achieve an anti-infective effect.

Salidroside Protects Against Influenza A Virus-Induced Acute Lung Injury in Mice.

Influenza A virus infections can cause acute lung injury (ALI) in humans; thus, the identification of potent antiviral agents is urgently required. Herein, the effects of salidroside on influenza A virus-induced ALI were investigated in a murine model. BALB/c mice were intranasally inoculated with H1N1 virus and treated with salidroside. The results of this study show that salidroside treatment (30 and 60 mg/kg) significantly attenuated the H1N1 virus-induced histological alterations in the lung and inhibited inflammatory cytokine production. Salidroside also decreased the wet/dry ratio, viral titers, and Toll-like receptor 4 expression in the lungs. Therefore, salidroside may represent a potential therapeutic reagent for the treatment of influenza A virus-induced ALI.

Analysis of Gut Microbiota in Patients with Coronary Artery Disease and Hypertension.

Cardiovascular and cerebrovascular diseases are characterized by high rates of morbidity and mortality. Microbiota is closely associated with cardiovascular disease. We aimed to comprehensively analyze the microbiotas of 300 healthy controls, 300 patients with high blood pressure (HBP), and 300 patients with coronary heart disease (CHD). The results indicated no significant difference in microbiota diversity among the three groups (P > 0.05). However, differences in microbiota richness among the three groups were significant (P < 0.05). Bacteroidetes and Bacteroidia were the dominant bacteria in the CHD group, Enterobacteriales and Escherichia-shigella in the HBP group, and Acidaminococcaceae and Phascolarctobacterium in the healthy control group. The prediction results of the random forest model indicated that the population with CHD displayed prominent features with high sensitivity, indicating that microbiota detection might become a novel clinical indicator to predict and monitor the risk of cardiovascular events. The prediction of microbiota function suggested differences in oxygen supply and chronic inflammation between populations with HBP/CHD and healthy populations. Although there is no difference in gut microbiota diversity among the three groups, each group has its dominant microbiota in terms of richness.

Measurement of the nerve root of the lower lumbar region using digital images.

The purpose of this study was to evaluate the relationship between the nerve root of lower lumbar and the surrounding structures using three-dimensional computed tomography (3D CT).Twenty-three consecutive patients with thoracolumbar fractures without obvious radiological degeneration were retrospectively studied at the spinal surgery department of the hospital. The parameters of the relationship between the nerve root of the lower lumbar and the surrounding structures were measured using 3D CT in the work station of the picture archiving and communication system.The size of the dorsal root ganglion (DRG) of the L4 was 5.5 ±â€Š0.4 mm on the right side and 5.8 ±â€Š0.3 mm on the left side. The size of the DRG of the L5 was 6.1 ±â€Š0.5 mm on the right side and 5.7 ±â€Š0.4 mm on the left side. The value of the preganglionic nerve root of the L4 was 11.2 ±â€Š0.6 mm on the right side and 12.3 ±â€Š0.8 mm on the left side, and the value of the preganglionic nerve root of the L5 was 15.1 ±â€Š1.1 mm on the right side and 14.9 ±â€Š0.9 mm on the left side.Using 3D CT imaging constructed in the picture archiving and communication system is a practical and convenient method for evaluating the relationship between the nerve root and the surrounding structures in the routine clinical work of a spinal surgeon. The data obtained through 3D CT imaging will be helpful for surgeons, allowing them to become more familiar with correlating anatomical knowledge of individual patient.

Qian Five Rhinoceros Gindeng (QFRG) protects against development of immune thrombocytopenia via miR-181a inhibition of TLR-4 expression.

AIMS: Traditional Chinese medicine (TCM) has been considered to be effective auxiliary strategy for the treatment of hemocytopenia including immune thrombocytopenia. However, the molecular mechanism is still not understood. METHODS: In present study, Qian Five Rhinoceros Gindeng (QFRG) mainly containing buffalo horn, rehmannia root, radix rubia, trogopterus dung and radix salviae miltiorrhizae administrated to thrombocytopenia mice induced by injection of MWReg30. MicroRNAs (miRNAs), Toll Like Receptors (TLRs) and cytokines were assayed in monocytes separated from mice peripheral blood. The relationship between miRNAs and TLRs was investigated in Mouse leukaemic monocyte macrophage cell line RAW264.7. RESULTS: The mice with administration of QFRG had a significant increase in platelet count, and miR-181a of monocytes was markedly up-regulated in QFRG treated group. QFRG also decreased the levels of TLR4, IL-6 and TNF-α. In addition, miR-181a inhibitor reversed the effects of QFRG on platelet count, TLR4 and cytokines. Overexpression of miR-181a in lipopolysaccharide-induced showed a decrease of TLR4, IL-6 and TNF-α level. CONCLUSIONS: QFRG protects against development of immune thrombocytopenia via miR-181a inhibition of TLR-4 expression.

Emodin prevents intima thickness via Wnt4/Dvl-1/ß-catenin signaling pathway mediated by miR-126 in balloon-injured carotid artery rats.

Neointimal proliferation after vascular injury is a key mechanism of restenosis, a major cause of percutaneous transluminal angioplasty failure and artery bypass occlusion. Emodin, an anthraquinone with multiple physiological activities, has been reported to inhibit proliferation of vascular smooth muscle cells (VSMCs) that might cause intimal arterial thickening. Thus, in this study, we established a rat model of balloon-injured carotid artery and investigated the therapeutic effect of emodin and its underlying mechanism. Intimal thickness was analyzed by hematoxylin and eosin staining. Expression of Wnt4, dvl-1, ß-catenin and collagen was determined by immunohistochemistry and/or western blotting. The proliferation of VSMC was evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and electron microscopy. MicroRNA levels were quantified by real-time quantitative PCR. Emodin relieved injury-induced artery intimal thickness. Results of western blots and immunohistochemistry showed that emodin suppressed expression of signaling molecules Wnt4/Dvl-1/ß-catenin as well as collagen protein in the injured artery. In addition, emodin enhanced expression of an artery injury-related microRNA, miR-126. In vitro, MTT assay showed that emodin suppressed angiotensin II (AngII)-induced proliferation of VSMCs. Emodin reversed AngII-induced activation of Wnt4/Dvl-1/ß-catenin signaling by increasing expression of miR-126 that was strongly supported by transfection of mimic or inhibitor for miR-126. Emodin prevents intimal thickening via Wnt4/Dvl-1/ß-catenin signaling pathway mediated by miR-126 in balloon-injured carotid artery of rats.

Wnt4/ß-catenin signaling pathway modulates balloon-injured carotid artery restenosis via disheveled-1.

BACKGROUND: Restenosis is a common adverse event of endovascular procedures and troubles cardiologists. However, the mechanism underlying restenosis is still not fully understood. To evaluate whether disheveled-1 (Dvl-1) is involved in the Wnt4/ß-catenin signaling pathway to participate in the mechanisms of vascular restenosis. METHODOLOGY: Rat model of balloon-injured carotid artery was established and atorvastatin was used to treat artery injury. Vascular smooth muscle cells (VSMC) were isolated from rats and cultured in DMEM exposed to AngII. Down-regulation and overexpression of Dvl-1 were conducted in cells to explore the role underlying its effects on VSMC proliferation and collagen expression. Adenovirus with overexpressing Dvl-1 was injected into rats to evaluate the role of Dvl-1 in artery injury rats. RESULTS: The results in vivo found that Wnt4, Dvl-1 and ß-catenin expression as well as collagen volume fraction (CVF) in injured artery were significantly increased. The results in vitro showed that Dvl-1 overexpression reversed the treatment effects of atorvastatin on VSMCs proliferation and collagen expression. It was also canceled by overexpressing Dvl-1 that the decrease of ß-catenin protein treated with atorvastatin in cells exposed to AngII. In addition, treated artery injury rats with atorvastatin, the group with injection of Ad-Dvl-1 had higher levels of intima thickness, intimal/medial area ratio and CVF. CONCLUSION: Dvl-1 was probably a key regulator in the pathway of wnt4/ß-catenin to take part in the vascular restenosis partly, and Dvl-1 is a potential gene to anti- restenosis.

[Effect of tetramethylpyrazine on the proliferation and collagen synthesis of vascular smooth muscle cells].

OBJECTIVE: To study the action mechanism of tetramethylpyrazine (TMP) on the proliferation of vascular smooth muscle cells (VSMCs), thus providing experimental evidence for Chinese medicine to effectively prevent restenosis. METHODS: Rats' thoracic aorta VSMCs in vitro cultured (cell line A7r5) were divided into five groups, i.e., the negative control group, the angiotensin II (Ang II, 10(-6) mol/L) group, the low dose TMP (20 micromol/L) plus Ang II group, the middle dose TMP (40 micromol/L) plus Ang II group, the high dose TMP (80 micromol/L) plus Ang II group. The proliferation ratio was detected by MTT. Gene and protein expressions of Wnt4, Dvl-1, beta-catenin, CyclinD1, and collagen I and III were detected with real-time fluorescent quantitative PCR and Western blot respectively. RESULTS: Compared with the negative control group, the proliferation ratio of VSMCs obviously increased in the Ang II group (P < 0.05). Compared with the Ang II group, the proliferation ratio of VSMCs obviously decreased in the middle dose TMP plus Ang II group and the high dose TMP plus Ang II group (P < 0.05). Compared with the negative control group, gene and protein expressions of Wnt4, Dvl-1, beta-catenin, CyclinD1, Col I, and Col III were obviously up-regulated in the Ang II group (P < 0.05). Compared with the Ang II group, mRNA and protein expressions of Wnt4, Dvl-1, beta-catenin, CyclinD1, Col I, and Col III were obviously down-regulated in the middle dose TMP plus Ang II group and the high dose TMP plus Ang II group (P < 0.05). The aforesaid indices were dose-dependent in the low, middle, and high dose TMP plus Ang II groups. CONCLUSION: TMP inhibited Ang II induced proliferation and collagen secretion of VSMCs through down-regulating Wnt signal pathway.

[Relationship between endothelial-to-mesenchymal transition and cardiac fibrosis in acute viral myocarditis].

OBJECTIVE: To investigate the relationship between endothelial-to-mesenchymal transition (EndMT) and myocardial fibrosis in acute viral myocarditis (VMC). METHODS: Twenty-eight Balb/c mice were randomized into 3 groups: control group (n=8), VMC group(n=10) and intervention group(n=10). Mice in VMC and intervention groups were injected intraperitoneally(i.p) with single dose of coxsackievirus B3, mice in control group were injected with equal amount of viral-free vehicle. In the following day, mice in control and VMC groups were injected i.p with 0.1 ml of saline and intervention group with 0.1 ml of recombinant human bone morphogenetic protein 7(rh-BMP7) at a concentration of 300 µg/kg. The mice hearts were harvested after 7 d, cardiac collagen volume fraction (CVF) was calculated on picrosirius red-stained sections. mRNA and protein expression of TGF-ß1, CD31, VE-cadherin, fibroblast special protein 1 (FSP-1) and α-smooth muscle actin (α-SMA) and collagen 1α1 in myocardiac tissues were detected by real-time RT-PCR and Western blot analysis, respectively. RESULTS: Compared to controls, overt fibrosis was presented in necrotic area of myocardium in VMC group. Meanwhile, marked increase of TGF-ß1 expression accompanied with EndMT characterized by loss of endothelial phenotype (decreased expression of CD31 and VE-cadherin), gain of mesenchymal proteins (overexpression of FSP-1 and α-SMA) and increased synthesis of collagen was also demonstrated. Both EndMT and cardiac fibrosis were simultaneously reversed by TGF-ß1 inhibition. CONCLUSION: EndMT is involved in cardiac fibrosis in acute viral myocarditis, TGF-ß1 might be a main mediator.